cell free system

Summary

Summary: A fractionated cell extract that maintains a biological function. A subcellular fraction isolated by ultracentrifugation or other separation techniques must first be isolated so that a process can be studied free from all of the complex side reactions that occur in a cell. The cell-free system is therefore widely used in cell biology. (From Alberts et al., Molecular Biology of the Cell, 2d ed, p166)

Top Publications

  1. ncbi Cell-free translation reconstituted with purified components
    Y Shimizu
    Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Bldg FSB401, Kasiwanoha, Kashiwa, Chiba Prefecture 277-8562, Japan
    Nat Biotechnol 19:751-5. 2001
  2. pmc Towards synthesis of a minimal cell
    Anthony C Forster
    Department of Pharmacology and Vanderbilt Institute of Chemical Biology, Vanderbilt University Medical Center, Nashville, TN 37232, USA
    Mol Syst Biol 2:45. 2006
  3. ncbi Bidirectional transport by distinct populations of COPI-coated vesicles
    L Orci
    Department of Morphology, Faculty of Medicine, University of Geneva Medical Center, Switzerland
    Cell 90:335-49. 1997
  4. ncbi Dual role of Rac in the assembly of NADPH oxidase, tethering to the membrane and activation of p67phox: a study based on mutagenesis of p67phox-Rac1 chimeras
    Rive Sarfstein
    Julius Friedrich Cohnheim Minerva Center for Phagocyte Research and the Ela Kodesz Institute of Host Defense against Infectious Diseases, Sackler School of Medicine, Tel Aviv University, Tel Aviv 69978, Israel
    J Biol Chem 279:16007-16. 2004
  5. ncbi In vitro conversion of full-length mammalian prion protein produces amyloid form with physical properties of PrP(Sc)
    Olga V Bocharova
    Medical Biotechnology Center, University of Maryland Biotechnology Institute, Baltimore, MD 21201, USA
    J Mol Biol 346:645-59. 2005
  6. ncbi Mimicking the Escherichia coli cytoplasmic environment activates long-lived and efficient cell-free protein synthesis
    Michael C Jewett
    Department of Chemical Engineering, Stanford University, Stanford, California 94305 5025, USA
    Biotechnol Bioeng 86:19-26. 2004
  7. ncbi Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome c
    X Liu
    Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia 30322, USA
    Cell 86:147-57. 1996
  8. ncbi The past, present and future of cell-free protein synthesis
    Federico Katzen
    Invitrogen Corporation, 1600 Faraday Avenue, Carlsbad, CA 92008, USA
    Trends Biotechnol 23:150-6. 2005
  9. pmc An integrated cell-free metabolic platform for protein production and synthetic biology
    Michael C Jewett
    Department of Chemical Engineering, Stanford University, Stanford, CA 94305 5025, USA
    Mol Syst Biol 4:220. 2008
  10. ncbi Binding of inositol phosphate to DNA-PK and stimulation of double-strand break repair
    L A Hanakahi
    Imperial Cancer Research Fund, South Mimms, Hertfordshire, United Kingdom
    Cell 102:721-9. 2000

Detail Information

Publications280 found, 100 shown here

  1. ncbi Cell-free translation reconstituted with purified components
    Y Shimizu
    Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Bldg FSB401, Kasiwanoha, Kashiwa, Chiba Prefecture 277-8562, Japan
    Nat Biotechnol 19:751-5. 2001
    ..Moreover, omission of a release factor allowed efficient incorporation of an unnatural amino acid using suppressor transfer RNA (tRNA)...
  2. pmc Towards synthesis of a minimal cell
    Anthony C Forster
    Department of Pharmacology and Vanderbilt Institute of Chemical Biology, Vanderbilt University Medical Center, Nashville, TN 37232, USA
    Mol Syst Biol 2:45. 2006
    ..Our proposed minimal genome is 113 kbp long and contains 151 genes. We detail building blocks already in place and major hurdles to overcome for completion...
  3. ncbi Bidirectional transport by distinct populations of COPI-coated vesicles
    L Orci
    Department of Morphology, Faculty of Medicine, University of Geneva Medical Center, Switzerland
    Cell 90:335-49. 1997
    ....
  4. ncbi Dual role of Rac in the assembly of NADPH oxidase, tethering to the membrane and activation of p67phox: a study based on mutagenesis of p67phox-Rac1 chimeras
    Rive Sarfstein
    Julius Friedrich Cohnheim Minerva Center for Phagocyte Research and the Ela Kodesz Institute of Host Defense against Infectious Diseases, Sackler School of Medicine, Tel Aviv University, Tel Aviv 69978, Israel
    J Biol Chem 279:16007-16. 2004
    ..This demonstrates that Rac has a dual role in the assembly of NADPH oxidase. One is to tether p67phox to the membrane; the other is to induce an "activating" conformational change in p67phox...
  5. ncbi In vitro conversion of full-length mammalian prion protein produces amyloid form with physical properties of PrP(Sc)
    Olga V Bocharova
    Medical Biotechnology Center, University of Maryland Biotechnology Institute, Baltimore, MD 21201, USA
    J Mol Biol 346:645-59. 2005
    ....
  6. ncbi Mimicking the Escherichia coli cytoplasmic environment activates long-lived and efficient cell-free protein synthesis
    Michael C Jewett
    Department of Chemical Engineering, Stanford University, Stanford, California 94305 5025, USA
    Biotechnol Bioeng 86:19-26. 2004
    ....
  7. ncbi Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome c
    X Liu
    Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia 30322, USA
    Cell 86:147-57. 1996
    ..Cells undergoing apoptosis in vivo showed increased release of cytochrome c to their cytosol, suggesting that mitochondria may function in apoptosis by releasing cytochrome c...
  8. ncbi The past, present and future of cell-free protein synthesis
    Federico Katzen
    Invitrogen Corporation, 1600 Faraday Avenue, Carlsbad, CA 92008, USA
    Trends Biotechnol 23:150-6. 2005
    ..We review the advances on this expanding technology and highlight the growing list of associated applications...
  9. pmc An integrated cell-free metabolic platform for protein production and synthetic biology
    Michael C Jewett
    Department of Chemical Engineering, Stanford University, Stanford, CA 94305 5025, USA
    Mol Syst Biol 4:220. 2008
    ....
  10. ncbi Binding of inositol phosphate to DNA-PK and stimulation of double-strand break repair
    L A Hanakahi
    Imperial Cancer Research Fund, South Mimms, Hertfordshire, United Kingdom
    Cell 102:721-9. 2000
    ..The involvement of inositol phosphate in DNA-PK-dependent NHEJ is of particular interest since the catalytic domain of DNA-PKcs is similar to that found in the phosphatidylinositol 3 (PI 3)-kinase family...
  11. ncbi Phosphorylation and activation of myosin by Rho-associated kinase (Rho-kinase)
    M Amano
    Division of Signal Transduction, Nara Institute of Science and Technology, 8916 5 Takayama, Ikoma 630 01
    J Biol Chem 271:20246-9. 1996
    ..The activated Rho-kinase subsequently phosphorylates MLC. This may partly account for the mechanism by which Rho regulates cytokinesis, cell motility, or smooth muscle contraction...
  12. ncbi Executioner caspase-3, -6, and -7 perform distinct, non-redundant roles during the demolition phase of apoptosis
    E A Slee
    Molecular Cell Biology Laboratory, Department of Genetics, The Smurfit Institute, Trinity College, Dublin 2, Ireland
    J Biol Chem 276:7320-6. 2001
    ....
  13. ncbi Two transmembrane aspartates in presenilin-1 required for presenilin endoproteolysis and gamma-secretase activity
    M S Wolfe
    Center for Neurologic Diseases, Harvard Medical School and Brigham and Women s Hospital, Boston, Massachusetts 02115, USA
    Nature 398:513-7. 1999
    ....
  14. pmc Uncoupling retro-translocation and degradation in the ER-associated degradation of a soluble protein
    Robert J Lee
    Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260, USA
    EMBO J 23:2206-15. 2004
    ..Moreover, addition of only the 19S cap of the proteasome catalyzed ATP-dependent export of the polypeptide substrate, which was degraded upon subsequent addition of the 20S particle...
  15. ncbi In vitro cell-free conversion of bacterial recombinant PrP to PrPres as a model for conversion
    Louise Kirby
    Institute for Animal Health, Compton Laboratories, Newbury, Berkshire RG20 7NN, UK
    J Gen Virol 84:1013-20. 2003
    ....
  16. ncbi Quantitative polysome analysis identifies limitations in bacterial cell-free protein synthesis
    Kelly A Underwood
    Biophysics Program, D118 Fairchild Science Building, Stanford, California 94305 5126, USA
    Biotechnol Bioeng 91:425-35. 2005
    ....
  17. ncbi The LuxS family of bacterial autoinducers: biosynthesis of a novel quorum-sensing signal molecule
    S Schauder
    Department of Molecular Biology, Princeton University, Princeton, NJ 08544 1014, USA
    Mol Microbiol 41:463-76. 2001
    ..This result suggests that, unlike quorum sensing via the family of related homoserine lactone autoinducers, AI-2 is a unique, 'universal' signal that could be used by a variety of bacteria for communication among and between species...
  18. pmc Bax directly induces release of cytochrome c from isolated mitochondria
    J M Jurgensmeier
    Program on Apoptosis and Cell Death Research, The Burnham Institute, 10901 North Torrey Pines Road, La Jolla, CA 92037, USA
    Proc Natl Acad Sci U S A 95:4997-5002. 1998
    ....
  19. pmc A vesicle bioreactor as a step toward an artificial cell assembly
    Vincent Noireaux
    Center for Studies in Physics and Biology, The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA
    Proc Natl Acad Sci U S A 101:17669-74. 2004
    ..The reactor can then sustain expression for up to 4 days with a protein production of 30 muM after 4 days. Oxygen diffusion and osmotic pressure are critical parameters to maintain expression and avoid vesicle burst...
  20. ncbi Cell-free synthesis of 15N-labeled proteins for NMR studies
    Kiyoshi Ozawa
    Research School of Chemistry, Australian National University, Canberra ACT, Australia
    IUBMB Life 57:615-22. 2005
    ..We review methodological aspects of cell-free in vitro protein synthesis based on an Escherichia coli cell extract, in particular with regard to the production of 15 N-labeled proteins for analysis by NMR spectroscopy...
  21. ncbi Semiautomated cell-free conversion of prion protein: applications for high-throughput screening of potential antiprion drugs
    Leonid Breydo
    Medical Biotechnology Center, University of Maryland Biotechnology Institute, Baltimore, MD 21201, USA
    Anal Biochem 339:165-73. 2005
    ..We thus propose that the conversion of rPrP to the amyloid isoform can serve as a high-throughput screen for possible inhibitors of PrP(Sc) formation and potential anti-TSE drugs...
  22. ncbi A single-step photolithographic interface for cell-free gene expression and active biochips
    Amnon Buxboim
    Departments of Materials and Interfaces, The Weizmann Institute of Science, PO Box 26, Rehovot 76100, Israel
    Small 3:500-10. 2007
    ..The Daisy molecule and described approach enables groups not proficient in surface chemistry to construct active biochips based on cell-free gene expression...
  23. ncbi Amino acid stabilization for cell-free protein synthesis by modification of the Escherichia coli genome
    Nathalie Michel-Reydellet
    Department of Chemical Engineering, Stanford University, CA 94305 5025, USA
    Metab Eng 6:197-203. 2004
    ..Cysteine, however, continues to be degraded. Cell-free protein synthesis with the modified cell extract produces increased yields of the cysteine-free protein Outer Membrane Protein T (OmpT)...
  24. ncbi High yield cell-free production of integral membrane proteins without refolding or detergents
    Jessica J Wuu
    Department of Chemical Engineering, Stanford University, Stanford, CA 94305, USA
    Biochim Biophys Acta 1778:1237-50. 2008
    ..Whereas TetA incorporates efficiently into vesicle membranes with over two-thirds of the synthesized protein being inserted, MtlA yields appear to be limited by insufficient concentrations of a membrane-associated chaperone...
  25. ncbi Substrate replenishment extends protein synthesis with an in vitro translation system designed to mimic the cytoplasm
    Michael C Jewett
    Department of Chemical Engineering, Stanford University, Stanford, California 94305 5025, USA
    Biotechnol Bioeng 87:465-72. 2004
    ..4 mg/mL of CAT. These results underscore the critical role that nucleotides play in the combined transcription-translation reaction and highlight the importance of understanding metabolic processes influencing substrate depletion...
  26. ncbi Protein synthesis in liposomes with a minimal set of enzymes
    Giovanni Murtas
    Centro Studi e Ricerche E Fermi, Compendio Viminale, 00184 Rome, Italy
    Biochem Biophys Res Commun 363:12-7. 2007
    ..The expression of GPAT and LPAAT in liposomes would in principle allow the production of the cell boundary from within...
  27. ncbi An economical method for cell-free protein synthesis using glucose and nucleoside monophosphates
    Kara A Calhoun
    Department of Chemical Engineering, Stanford University, Stanford, CA 94305 5025, USA
    Biotechnol Prog 21:1146-53. 2005
    ..The glucose/NMP cell-free reaction system dramatically reduces reagent costs while supplying high protein yields...
  28. pmc Thioredoxin is required for vacuole inheritance in Saccharomyces cerevisiae
    Z Xu
    Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire, 03755 3844, USA
    J Cell Biol 132:787-94. 1996
    ..A deletion of both S. cerevisiae thioredoxin genes causes a striking vacuole inheritance defect in vivo, establishing a role for thioredoxin as a novel factor in this trafficking reaction...
  29. ncbi Incorporation of nonnatural amino acids into proteins by using various four-base codons in an Escherichia coli in vitro translation system
    T Hohsaka
    Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University, 3-1-1 Tsushimanaka, Okayama 700-8530, Japan
    Biochemistry 40:11060-4. 2001
    ..This work shows that the four-base codon strategy is more advantageous than the amber suppression strategy in efficiency and versatility...
  30. pmc GTP-binding Ypt1 protein and Ca2+ function independently in a cell-free protein transport reaction
    D Baker
    Division of Biochemistry and Molecular Biology, University of California, Berkeley 94720
    Proc Natl Acad Sci U S A 87:355-9. 1990
    ..Thus, Ypt1p and Ca2+ are required at distinct steps...
  31. ncbi Efficient protein selection based on ribosome display system with purified components
    Hiroyuki Ohashi
    The Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, FSB 401, 5 1 5, Kashiwanoha, Kashiwa, Chiba 277 8562, Japan
    Biochem Biophys Res Commun 352:270-6. 2007
    ..Urabe, T. Yomo, Nascent chain, RNA, and ribosome complexes generated by pure translation system (see the accompanying paper).], demonstrate that the PURE system can provide a basis for reliable and reproducible ribosome display...
  32. pmc A cell-free protein synthesis system for high-throughput proteomics
    Tatsuya Sawasaki
    Department of Applied Chemistry, Faculty of Engineering, and Venture Business Laboratory, Ehime University, Matsuyama 790 8577, Japan
    Proc Natl Acad Sci U S A 99:14652-7. 2002
    ..The cell-free system, reported here, bypasses many of the time-consuming cloning steps of conventional expression systems and lends itself to a robotic automation for the high-throughput expression of proteins...
  33. ncbi Self-assembling protein microarrays
    Niroshan Ramachandran
    Harvard Institute of Proteomics, Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 320 Charles Street, Cambridge, MA 02141, USA
    Science 305:86-90. 2004
    ..We used the technology to map pairwise interactions among 29 human DNA replication initiation proteins, recapitulate the regulation of Cdt1 binding to select replication proteins, and map its geminin-binding domain...
  34. ncbi The mRNA-binding protein YB-1 (p50) prevents association of the eukaryotic initiation factor eIF4G with mRNA and inhibits protein synthesis at the initiation stage
    Maxim P Nekrasov
    Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region, 142290 Russian Federation, Russia
    J Biol Chem 278:13936-43. 2003
    ..The accumulation is accompanied by eukaryotic initiation factor eIF4G dissociation from mRNA. The C-terminal domain of YB-1 is responsible for inhibition of translation as well as the disruption of mRNA interaction with eIF4G...
  35. ncbi Genome-scale, biochemical annotation method based on the wheat germ cell-free protein synthesis system
    Tatsuya Sawasaki
    Cell Free Science and Technology Research Center, The Venture Business Laboratory, Ehime University, Matsuyama 790 8577, Japan
    Phytochemistry 65:1549-55. 2004
    ..The information on biochemical function of gene products accumulated should revolutionize our understanding of biology and fundamentally alter the practice of medicine and influence other industries as well...
  36. ncbi Generation of high density protein microarrays by cell-free in situ expression of unpurified PCR products
    Philipp Angenendt
    Functional Genome Analysis, German Cancer Research Center, Im Neuenheimer Feld 580, D 69120 Heidelberg, Germany
    Mol Cell Proteomics 5:1658-66. 2006
    ..Finally we adapted the system to the high throughput expression of libraries by designing a single primer pair for the introduction of the required T7 promoter and demonstrated the in situ expression using 384 randomly chosen clones...
  37. pmc Principles of cell-free genetic circuit assembly
    Vincent Noireaux
    Center for Studies in Physics and Biology, The Rockefeller University, New York, NY 10021, USA
    Proc Natl Acad Sci U S A 100:12672-7. 2003
    ..Faster turnover of RNA message can relieve competition between genes and stabilize output against variations in input and parameters...
  38. ncbi Pure translation display
    Anthony C Forster
    Department of Pathology, Brigham and Women s Hospital, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02115, USA
    Anal Biochem 333:358-64. 2004
    ..Pure translation display should enable the directed evolution of peptide analogs with desirable catalytic or pharmacological properties...
  39. pmc Continued protein synthesis at low [ATP] and [GTP] enables cell adaptation during energy limitation
    Michael C Jewett
    Department of Chemical Engineering, Stanford University, Stanford, CA 94305 5025, USA
    J Bacteriol 191:1083-91. 2009
    ..We anticipate that cell-free studies in which complex metabolic systems are activated will be valuable tools for elucidating the behavior of such systems...
  40. ncbi Methods for high-throughput materialization of genetic information based on wheat germ cell-free expression system
    Tatsuya Sawasaki
    Cell Free Science and Technology Research Center, Ehime University, Matsuyama, Japan
    Methods Mol Biol 375:95-106. 2007
    ..Here, we describe protocols for this cell-free expression system...
  41. ncbi Ca2+/calmodulin signals the completion of docking and triggers a late step of vacuole fusion
    C Peters
    Friedrich Miescher Laboratorium der Max Planck Gesellschaft, Tubingen, Germany
    Nature 396:575-80. 1998
    ..However, the origin of the Ca2+ signal, its receptor and its mode of processing differ...
  42. ncbi Production of membrane proteins using cell-free expression systems
    Lavinia Liguori
    University Joseph Fourier, HumProTher Laboratory, GREPI, CHU Grenoble, 38043 Grenoble, France
    Expert Rev Proteomics 4:79-90. 2007
    ..In this review, we highlight the recent improvements of these cell-free expression systems and their direct applications in the fields of membrane proteins production, protein therapy and modern proteomics...
  43. pmc Tyrosine, cysteine, and S-adenosyl methionine stimulate in vitro [FeFe] hydrogenase activation
    Jon M Kuchenreuther
    Department of Chemical Engineering, Stanford University, Stanford, California, United States of America
    PLoS ONE 4:e7565. 2009
    ..This unique [6Fe-6S] complex contains multiple non-protein moieties and requires several maturation enzymes for its assembly. The pathways and biochemical precursors for H-cluster biosynthesis have yet to be elucidated...
  44. ncbi A novel cell-free protein synthesis system
    Kalavathy Sitaraman
    SAIC National Cancer Institute at Frederick, 1050 Boyles Street, Building 327, Frederick, MD 21702 1201, USA
    J Biotechnol 110:257-63. 2004
    ..Therefore, combination of these improvements should be amenable to rapid expression of proteins in a high-throughput manner for proteomics and structural genomics applications...
  45. ncbi Diacylglycerol induces fusion of nuclear envelope membrane precursor vesicles
    Teresa Barona
    Biology Department, Amherst College, Amherst, Massachusetts 01002, USA
    J Biol Chem 280:41171-7. 2005
    ....
  46. ncbi Automated system for high-throughput protein production using the dialysis cell-free method
    Masaaki Aoki
    RIKEN Systems and Structural Biology Center SSBC, 1 7 22 Suehiro cho, Tsurumi ku, Yokohama 230 0045, Japan
    Protein Expr Purif 68:128-36. 2009
    ....
  47. ncbi Cell-free synthesis of a functional ion channel in the absence of a membrane and in the presence of detergent
    Catherine Berrier
    Institut de Biochimie et de Biophysique Moleculaire et Cellulaire, Unite Mixte de Recherche CNRS 8619, Bat 430, Universite Paris Sud, 91405 Orsay Cedex, France
    Biochemistry 43:12585-91. 2004
    ..Cell-free synthesis of membrane proteins should prove a valuable tool for the production of membrane proteins whose overexpression in heterologous systems is difficult...
  48. ncbi Development of cell-free protein synthesis platforms for disulfide bonded proteins
    Aaron R Goerke
    Department of Chemical Engineering, Stanford University, Stanford, California 94305, USA
    Biotechnol Bioeng 99:351-67. 2008
    ..Establishing general CFPS platforms enhances the potential for cell-free protein synthesis to reliably produce complex protein products at low production and capital costs with very rapid process development timelines...
  49. ncbi High level cell-free expression and specific labeling of integral membrane proteins
    Christian Klammt
    Centre for Biomolecular Magnetic Resonance, University of Frankfurt Main, Institute for Biophysical Chemistry, Frankfurt Main, Germany
    Eur J Biochem 271:568-80. 2004
    ....
  50. ncbi Cell-free protein synthesis for proteomics
    Alison M Jackson
    Discerna Ltd, Babraham Research Campus, Babraham, Cambridge CB2 4AT, UK
    Brief Funct Genomic Proteomic 2:308-19. 2004
    ..This paper reviews the most commonly used cell-free systems and their applications in proteomics...
  51. ncbi Expression of G protein coupled receptors in a cell-free translational system using detergents and thioredoxin-fusion vectors
    Goshi Ishihara
    RIKEN Genomic Sciences Center, 1 7 22 Suehiro cho, Tsurumi, Yokohama 230 0045, Japan
    Protein Expr Purif 41:27-37. 2005
    ..This cell-free translation system (about 1mg/ml of reaction volume for 6-8 h) has biophysical and biochemical advantages for the synthesis of integral membrane proteins...
  52. ncbi Effect of energy source on the efficiency of translational termination during cell-free protein synthesis
    Jin Ho Ahn
    Interdisciplinary Program for Biochemical Engineering and Biotechnology, College of Engineering, Seoul National University, Seoul 151 742, Republic of Korea
    Biochem Biophys Res Commun 337:325-9. 2005
    ....
  53. ncbi Microbead display of proteins by cell-free expression of anchored DNA
    Olof Nord
    Department of Biotechnology, Royal Institute of Technology KTH, AlbaNova University Center, SE 106 91 Stockholm, Sweden
    J Biotechnol 106:1-13. 2003
    ..The use of this technology, including alternative formats, for different proteomics applications is discussed...
  54. ncbi Protein chip fabrication by capture of nascent polypeptides
    Sheng Ce Tao
    Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA
    Nat Biotechnol 24:1253-4. 2006
    ..Using these approaches, we efficiently fabricated both peptide and protein microarrays at relatively high density. We further demonstrated that such protein chips can be used to analyze protein activity...
  55. ncbi Toward an artificial cell based on gene expression in vesicles
    Vincent Noireaux
    Center for Studies in Physics and Biology, The Rockefeller University, New York, NY 10021, USA
    Phys Biol 2:P1-8. 2005
    ..The specific degradation complex ClpXP from E. coli is introduced to create a sink for the synthesized proteins. Perspectives and subsequent limitations of this approach are discussed...
  56. ncbi Chaperone-assisted folding of a single-chain antibody in a reconstituted translation system
    Bei Wen Ying
    Department of Medical Genome Sciences, Graduate School of Frontier Sciences, Kashiwa, Chiba, Japan
    Biochem Biophys Res Commun 320:1359-64. 2004
    ..In addition, both systems also acted as chaperones after translation had been stopped. In contrast, the GroEL/ES system showed little or no co- or post-translational assistance in folding...
  57. ncbi Structural genomics projects in Japan
    S Yokoyama
    Genomic Sciences Center, RIKEN Yokohama Institute, Tsurumi, Japan
    Nat Struct Biol 7:943-5. 2000
    ..The newest, the structural genomics project at the Biological Information Research Center, focuses on human membrane proteins...
  58. ncbi Expression of a cascading genetic network within liposomes
    Keitaro Ishikawa
    Department of Biotechnology, Graduate School of Engineering, Osaka University, 2 1 Yamadaoka, Suita, Osaka 565 0871, Japan
    FEBS Lett 576:387-90. 2004
    ..We show that the two-stage genetic network constructed in a cell-free expression system is functional within liposomes...
  59. ncbi High-throughput cell-free systems for synthesis of functionally active proteins
    Alexander S Spirin
    Institute of Protein Research, Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russia
    Trends Biotechnol 22:538-45. 2004
    ....
  60. pmc Complexes between the LKB1 tumor suppressor, STRAD alpha/beta and MO25 alpha/beta are upstream kinases in the AMP-activated protein kinase cascade
    Simon A Hawley
    Division of Molecular Physiology, University of Dundee, Dundee DD1 5EH, UK
    J Biol 2:28. 2003
    ..We recently showed that LKB1 exists as a complex with two accessory subunits, STRAD alpha/beta and MO25 alpha/beta...
  61. ncbi A bilayer cell-free protein synthesis system for high-throughput screening of gene products
    Tatsuya Sawasaki
    Department of Applied Chemistry, Faculty of Engineering, The Venture Business Laboratory, Ehime University, 790 8577, Matsuyama, Japan
    FEBS Lett 514:102-5. 2002
    ..This novel bilayer-based cell-free protein synthesis system with its simplicity, minimum time and low cost may be useful practical methodology in the post-genome era...
  62. ncbi Exploration of the diaphorase activity of neutrophil NADPH oxidase
    Alexandra Poinas
    Laboratoire de Biochimie et Biophysique des Systemes Integres, UMR 5092 CEA CNRS, D├ępartement de Biologie Mol├ęculaire et Structurale Grenoble, France
    Eur J Biochem 269:1243-52. 2002
    ..Finally, the heme inhibitor bipyridyl competitively inhibited the reduction of INT in anaerobiosis, and the oxygen uptake in aerobiosis...
  63. ncbi An unnatural base pair for incorporating amino acid analogs into proteins
    Ichiro Hirao
    Yokoyama CytoLogic Project, ERATO, JST, c o RIKEN, 2 1 Hirosawa, Wako Shi, Saitama 351 0198, Japan
    Nat Biotechnol 20:177-82. 2002
    ..This coupled transcription-translation system will permit the efficient synthesis of proteins with a tyrosine analog at the desired position...
  64. ncbi Rapid production of milligram quantities of proteins in a batch cell-free protein synthesis system
    Tae Wan Kim
    School of Chemical and Biological Engineering, College of Engineering, Seoul National University, Korea
    J Biotechnol 124:373-80. 2006
    ..The simple, fast, and highly productive cell-free protein synthesis system described herein should offer a versatile platform for the preparation of protein molecules in various post-genomic efforts...
  65. ncbi [High-throughput expression of proteins from cDNAs catalogue from Arabidopsis in wheat germ cell-free protein synthesis system]
    Tatsuya Sawasaki
    Tanpakushitsu Kakusan Koso 47:1003-8. 2002
  66. ncbi A role for G1/S cyclin-dependent protein kinases in the apoptotic response to ionizing radiation
    Carla V Finkielstein
    Howard Hughes Medical Institute and Department of Pharmacology, University of Colorado School of Medicine, Denver, Colorado 80262, USA
    J Biol Chem 277:38476-85. 2002
    ..These data suggest that caspase cleavage of both cyclin D1-Cdk4 and cyclin A2-Cdk2 promotes specific apoptotic events in embryos undergoing apoptosis in response to ionizing radiation...
  67. ncbi Alpha-crystallin promotes assembly of a trimeric form of Mycobacterium tuberculosis Hsp16.3 in a cell free system
    A Abulimiti
    Department of Biological Science and Biotechnology, School of Life Science, Tsinghua University, The Protein Science Laboratory of the Education Ministry, Beijing, 100084 P R China
    Biochemistry (Mosc) 68:269-74. 2003
    ..3, was also detected in the in vitro expression system. A latent phase of ~10 min in the appearance of the first detectable species indicated that Hsp16.3 assembly did not occur co-translationally...
  68. ncbi Dissecting the mechanism and assembly of a complex virulence mycobacterial lipid
    Omita A Trivedi
    National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi 110 067, India
    Mol Cell 17:631-43. 2005
    ..Apart from elucidating mechanisms that generate chemical heterogeneity with PDIMs, this study also presents an attractive approach to explore host-pathogen interactions by altering mycobacterial surface coat...
  69. ncbi Expression of soluble recombinant proteins in a cell-free system using a 96-well format
    Didier Busso
    Department of Chemistry, University of California, Berkeley, 314 Calvin Lab MS5230, CA 94720 5230, USA
    J Biochem Biophys Methods 55:233-40. 2003
    ..The identification of soluble proteins is performed by the Dot Blot procedure using an anti-His tag antibody. The applications and the automation of this method are described...
  70. pmc Nuclear envelope assembly is promoted by phosphoinositide-specific phospholipase C with selective recruitment of phosphatidylinositol-enriched membranes
    Richard D Byrne
    Cell Biophysics Laboratory, London Research Institute LRI, Cancer Research UK CR UK, 44, Lincoln s Inn Fields, London, WC2A 3PX, UK
    Biochem J 387:393-400. 2005
    ..We have exploited this and the intervention studies to illustrate a pivotal role for PI-PLC and its product DAG in the formation of NEs...
  71. ncbi Effect of poly(ADP-ribose) polymerase inhibitors on oxidative stress evoked hydroxyl radical level and macromolecules oxidation in cell free system of rat brain cortex
    Grzegorz A Czapski
    Medical Research Centre, Polish Academy of Sciences, 5 Pawinskiego St, PL 02106 Warsaw, Poland
    Neurosci Lett 356:45-8. 2004
    ..These results indicate that 3-A beta, benzamide and DHIQ are potent hydroxyl radical scavengers and antioxidants. These data ought to be taken into consideration when properties of these compounds as PARP inhibitors are evaluated...
  72. ncbi Effective non-viral leader for cap-independent translation in a eukaryotic cell-free system
    L A Shaloiko
    Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region, Russia 142290
    Biotechnol Bioeng 88:730-9. 2004
    ..We propose the obelin mRNA leader be used for effective cap-independent translation in eukaryotic cell-free systems, including combined transcription-translation systems with uncontrolled phage polymerase-catalyzed accumulation of mRNA...
  73. ncbi Formation of circular polyribosomes in wheat germ cell-free protein synthesis system
    Kairat Madin
    Cell Free Science and Technology Research Center, and The Venture Business Laboratory, Ehime University, Matsuyama 790 8577, Japan
    FEBS Lett 562:155-9. 2004
    ..The wheat germ cell-free system may provide a good experimental system for understanding functional ribosomes at the molecular level...
  74. ncbi A novel way of amino acid-specific assignment in (1)H-(15)N HSQC spectra with a wheat germ cell-free protein synthesis system
    Eugene Hayato Morita
    Division of Gene Research, Department of Molecular Science, Integrated Center for Science, Ehime University, 3 5 7 Tarumi, Ehime 790 8566, Japan
    J Biomol NMR 30:37-45. 2004
    ..This is also the first direct experimental evidence of the presence of active transaminases and glutamine synthase in wheat germ extracts...
  75. ncbi Rapid expression of vaccine proteins for B-cell lymphoma in a cell-free system
    Junhao Yang
    Department of Chemical Engineering, Stanford University, Stanford, California 94305, USA
    Biotechnol Bioeng 89:503-11. 2005
    ..Between the two amino-terminal GM-CSF fusion proteins, GM-VL-VH showed a higher total and soluble yield than GM-VH-VL...
  76. ncbi Bacterial cell-free system for high-throughput protein expression and a comparative analysis of Escherichia coli cell-free and whole cell expression systems
    T V S Murthy
    Harvard Institute of Proteomics, 320 Charles Street, Cambridge, MA 02141, USA
    Protein Expr Purif 36:217-25. 2004
    ....
  77. ncbi Energizing cell-free protein synthesis with glucose metabolism
    Kara A Calhoun
    Department of Chemical Engineering, Stanford University, Stanford, California 94305 5025, USA
    Biotechnol Bioeng 90:606-13. 2005
    ....
  78. ncbi High-throughput, genome-scale protein production method based on the wheat germ cell-free expression system
    Yaeta Endo
    Department of Applied Chemistry, Faculty of Engineering, Ehime University, Matsuyama790 8577, Japan
    Biotechnol Adv 21:695-713. 2003
    ..The cell-free system described here bypasses most of the biological processes and lends itself to robotic automation for high-throughput expression of genetic information, thus opening up many possibilities in the post-genome era...
  79. ncbi Rapid expression of functional genomic libraries
    Kim A Woodrow
    Department of Chemical Engineering, Stanford University, Stanford, California 94305 5025, USA
    J Proteome Res 5:3288-300. 2006
    ..These improvements in the parallel synthesis of linear ETs combined with enhanced in vitro enzyme activation help to make CFPS systems more attractive platforms for high-throughput evaluation of protein function...
  80. ncbi The wheat germ cell-free expression system: methods for high-throughput materialization of genetic information
    Tatsuya Sawasaki
    Cell Free Science and Technology Research Center, Ehime University, Matsuyama, Japan
    Methods Mol Biol 310:131-44. 2005
    ..This chapter contains protocols for high-throughput protein production based on the cell-free system prepared from eukaryote wheat embryos...
  81. ncbi Production of protein for nuclear magnetic resonance study using the wheat germ cell-free system
    Toshiyuki Kohno
    Molecular Structure Research Group, Mitsubishi Kagaku Institute of Life Sciences MITILS, Tokyo, Japan
    Methods Mol Biol 375:257-72. 2007
    ..Here, we describe the method to produce proteins for NMR experiments using the improved wheat germ cell-free system, which exhibits several attractive features for high-throughput NMR study of proteins...
  82. ncbi Simultaneous expression and maturation of the iron-sulfur protein ferredoxin in a cell-free system
    Marcus E Boyer
    Department of Chemical Engineering, Stanford University, California 94305, USA
    Biotechnol Bioeng 94:128-38. 2006
    ..To our knowledge, this is the first demonstration of directed, high-yield production and maturation of an Fe-S protein in a cell-free system...
  83. pmc Targeted mRNA degradation by double-stranded RNA in vitro
    T Tuschl
    The Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA
    Genes Dev 13:3191-7. 1999
    ..Furthermore, preincubation of dsRNA potentiates its activity. These results demonstrate that RNAi can be mediated by sequence-specific processes in soluble reactions...
  84. ncbi Cyclin/Cdk-dependent initiation of DNA replication in a human cell-free system
    T Krude
    Wellcome Trust Cancer Research Campaign Institute, University of Cambridge, United Kingdom
    Cell 88:109-19. 1997
    ..In S phase cytosol, cyclins A/Cdk2 and E/Cdk2 triggered initiation synergistically...
  85. ncbi Cell-free synthesis and maturation of [FeFe] hydrogenases
    Marcus E Boyer
    Department of Chemical Engineering, Stanford University, 381 North South Mall, Stanford, California 94305, USA
    Biotechnol Bioeng 99:59-67. 2008
    ....
  86. ncbi Cell-free expression systems for eukaryotic protein production
    Yaeta Endo
    Cell Free Science and Technology Research Center, Ehime University, Matsuyama, Japan
    Curr Opin Biotechnol 17:373-80. 2006
    ..Several advances in the use of cell-free expression systems have been made in the past few years and successful applications of these systems to produce proteins for functional and structural biology studies have been reported...
  87. pmc Cdc6 protein causes premature entry into S phase in a mammalian cell-free system
    K Stoeber
    Wellcome CRC Institute, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK
    EMBO J 17:7219-29. 1998
    ..Addition of recombinant Cdc6 to permeabilized, but not intact, G1 nuclei causes up to 82% of the nuclei to initiate and accelerates G1 progression, making nuclei competent to replicate prematurely...
  88. pmc DNA end-joining catalyzed by human cell-free extracts
    P Baumann
    Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, Herts EN6 3LD, United Kingdom
    Proc Natl Acad Sci U S A 95:14066-70. 1998
    ....
  89. ncbi Wheat germ cell-free platform for eukaryotic protein production
    Dmitriy A Vinarov
    Center for Eukaryotic Structural Genomics, Biochemistry Department, University of Wisconsin Madison, Madison, WI, USA
    FEBS J 273:4160-9. 2006
    ..We present protein yields and costs and compare the wheat germ cell-free approach with alternative methods. Finally, we discuss remaining bottlenecks and approaches to their solution...
  90. pmc Cell-free synthetic biology: a bottom-up approach to discovery by design
    Michael L Simpson
    Center for Nanophase Materials Sciences, Oak Ridge National Laboratory, Oak Ridge, TN, USA
    Mol Syst Biol 2:69. 2006
  91. pmc Solid-phase translation and RNA-protein fusion: a novel approach for folding quality control and direct immobilization of proteins using anchored mRNA
    Manish Biyani
    Rational Evolutionary Design of Advanced Biomolecules, Saitama Small Enterprise Promotion Corporation SKIP City, 3 12 18 Kamiaoki, Kawaguchi, Saitama 333 0844, Japan
    Nucleic Acids Res 34:e140. 2006
    ....
  92. ncbi Synergistic and strain-specific effects of bovine spongiform encephalopathy and scrapie prions in the cell-free conversion of recombinant prion protein
    Martin Eiden
    Institute for Novel and Emerging Infectious Diseases at the Friedrich Loeffler Institut, Federal Research Institute for Animal Health, Boddenblick 5a, 17493 Greifswald Insel Riems, Germany
    J Gen Virol 87:3753-61. 2006
    ..This result indicates a synergistic effect of both strains during cell-free conversion. It is not yet known whether such a cooperative action between BSE and scrapie prions also occurs in vivo...
  93. ncbi Simple procedures for the construction of a robust and cost-effective cell-free protein synthesis system
    Tae Wan Kim
    School of Chemical and Biological Engineering, College of Engineering, Seoul National University, Seoul 151 742, Republic of Korea
    J Biotechnol 126:554-61. 2006
    ..Compared to the present standard procedures for the preparation of the S30 extract, the overall cost of the reagents and processing time were reduced by 80 and 60%, respectively...
  94. pmc The checkpoint protein MAD2 and the mitotic regulator CDC20 form a ternary complex with the anaphase-promoting complex to control anaphase initiation
    G Fang
    Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA
    Genes Dev 12:1871-83. 1998
    ..We conclude that the checkpoint-mediated cell cycle arrest involves hMAD2 receiving an upstream signal to inhibit activation of APC...
  95. pmc Translation of non-capped mRNAs in a eukaryotic cell-free system: acceleration of initiation rate in the course of polysome formation
    Olga M Alekhina
    Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region, Russia
    Nucleic Acids Res 35:6547-59. 2007
    ..The eIF4F-mediated circularization of polysomes may be considered as a possible event that leads to the re-initiation switch and the resultant acceleration effect...
  96. ncbi Rapid discovery of protein interactions by cell-free protein technologies
    M He
    Technology Research Group, The Babraham Institute, Cambridge CB22 3AT, U K
    Biochem Soc Trans 35:962-5. 2007
    ..These technologies can be combined for rapid detection of protein interactions...
  97. ncbi Functional protein expression from a DNA based wheat germ cell-free system
    Kate Qin Zhao
    Promega Corporation, 2800 Woods Hollow Road, Madison, WI 53711, USA
    J Struct Funct Genomics 8:199-208. 2007
    ..Thus, this efficient cell-free expression system can facilitate both functional and structural proteomics...
  98. ncbi Reversibility of scrapie-associated prion protein aggregation
    M A Callahan
    Rocky Mountain Laboratories, NIAID, National Institutes of Health, Hamilton, Montana 59840, USA
    J Biol Chem 276:28022-8. 2001
    ..Overall, the results suggest that PrP aggregation is not appreciably reversible under physiological conditions, but dissociation and refolding can be enhanced by treatments with GdnHCl...
  99. ncbi Regeneration of adenosine triphosphate from glycolytic intermediates for cell-free protein synthesis
    D M Kim
    Department of Chemical Engineering, Stanford University, Stanford, California 94305-5025, USA
    Biotechnol Bioeng 74:309-16. 2001
    ..As a result, the methods described provide cell-free protein synthesis with greater flexibility and cost efficiency...
  100. pmc A novel cell-free system reveals a mechanism of circular DNA formation from tandem repeats
    S Cohen
    Institute of Human Genetics, CNRS, Genome Dynamics and Development, 141 rue de la Cardonille, 34396 Montpellier Cedex 5, France
    Nucleic Acids Res 29:2542-8. 2001
    ....
  101. pmc The Sec13p complex and reconstitution of vesicle budding from the ER with purified cytosolic proteins
    N R Salama
    Department of Molecular and Cell Biology, Howard Hughes Research Institute, University of California, Berkeley 94720
    EMBO J 12:4073-82. 1993
    ..The vesicles formed with the purified protein fractions are competent to fuse with the Golgi and are biochemically distinct from the ER membrane fraction from which they derive...

Research Grants64

  1. The Role of PtdIns-5-P in Cell Function and Signaling
    LUCIA RAMEH-PLANT; Fiscal Year: 2012
    ..Our aim is to understand the function of this lipid under physiological and pathological conditions, with the goal of designing new strategies to prevent and treat diseases such as diabetes and bacterial infections. ..
  2. Role of Ncb5or in Insulin Production
    Hao Zhu; Fiscal Year: 2010
    ..These studies may provide new insights into the pathogenesis and treatment of diabetes. ..
  3. Host-pathogen signaling in group A streptococcal infection
    Michael R Wessels; Fiscal Year: 2012
    ..Understanding this process may lead to new strategies for prevention or treatment of GAS infection. ..
  4. Micro-RNA 122 and Chronic Hepatitis C
    Stanley M Lemon; Fiscal Year: 2013
    ..The studies proposed will provide novel insight into how it functions in HCV replication and in the pathogenesis of chronic hepatitis C, thereby enhancing the scientific background on which it can be exploited as a therapeutic target. ..
  5. Temporal and Spatial Signaling in Chemotaxis
    Peter N Devreotes; Fiscal Year: 2013
    ..abstract_text> ..
  6. Epigenetic regulation of transcription by mixed lineage leukemia protein MLL1
    Yali Dou; Fiscal Year: 2013
    ..This project is to understand the fundamental mechanism for these changes and thus, the cause of the disease. It will set up the foundation for potential medical interference in the future. ..
  7. Functional role of a host metabolic enzyme in viral replication
    Peter Nagy; Fiscal Year: 2010
    ..The proposed work will lead to deeper understanding of the role of the host in RNA virus infections and likely open new antiviral strategies. ..
  8. Secretion, protein targeting, and lipid metabolism in African Trypanosomes
    James D Bangs; Fiscal Year: 2013
    ..These studies will broaden our knowledge of basic cell biological processes in all eukaryotes, and will lay the groundwork for targeting the sphingolipid pathway for novel drug development. ..
  9. Novel Approaches to Access GPIs and GPI-Anchored Proteins for the Study of GPI An
    Zhongwu Guo; Fiscal Year: 2012
    ....
  10. Biochemical Dissection of the RIG-I Antiviral Pathway
    Zhijian J Chen; Fiscal Year: 2013
    ..If successful, our research should significantly advance the fields of immunology and virology, and provide new approaches to harness the host innate immunity to combat viral and autoimmune diseases. ..
  11. Regulation of Sindbis Virus Subgenomic RNA Synthesis
    Victor Stollar; Fiscal Year: 2010
    ..What we learn about the synthesis of SV SG RNA should have application to other viruses which make SG RNAs. As well, this new information could facilitate the development of SG RNA synthesis as a target for anti-viral therapy. ..
  12. The Fanconi anemia pathway: role in DNA interstrand cross-link repair
    Johannes Walter; Fiscal Year: 2013
    ..We have discovered that extracts derived from frog eggs recapitulate the process of ICL repair in a cell-free environment. We will use this system to elucidate the function of FA proteins in ICL repair. ..
  13. RHODOPSIN TRAFFICKING AND PHOTORECEPTOR MEMBRANE RENEWAL
    DUSANKA DANIELLE DERETIC; Fiscal Year: 2013
    ..This is expected to provide the foundation for improved therapeutic strategies for treatment of retina-specific and syndromic forms of photoreceptor loss. ..
  14. DNA Damage Checkpoint Recovery and Cancer
    Aimin Peng; Fiscal Year: 2013
    ..We will then directly evaluate new therapeutic ideas in which targeting Gwl or its initial activation during checkpoint recovery leads to more effective and specific cancer therapy. ..
  15. Mechanisms of Stress Induced Apoptosis in T-Cells
    Douglas R Green; Fiscal Year: 2011
    ..In this integrated approach to the problem of p53 function in apoptosis, we will elucidate the roles of p53 in the context of stress-induced apoptosis in mature and developing T cells. ..
  16. Early Events in Lipoprotein Assembly
    CHRISTOPHER JAMES MCKNIGHT; Fiscal Year: 2010
    ..The specific aims are: 1) To express N-terminal fragments of ApoS in a cell free system supplemented with phospholipids and phospholipid-triacylglycerol emulsions to test the hypothesis that ..
  17. Identifying the molecular basis of the maternal to zygotic transition
    Aaron F Straight; Fiscal Year: 2013
    ..Using this cell free system we have shown that adding DNA-coated beads to extracts lowers the threshold DNA concentration for ..
  18. The role of proprotein convertases in lipid metabolism
    Weijun Jin; Fiscal Year: 2009
    ....
  19. Role of Cytokine Signaling in Intestinal Restitution
    Narendra Kumar; Fiscal Year: 2013
    ..To test the hypothesis the specific objectives of the present proposal are: (a) to determine in a cell free system the specificity of Jak3 interactions with villin and ShcA and its significance on the regulation of Jak3-..
  20. Antiviral Mechanisms of 2 5A Dependent RNase L
    Robert H Silverman; Fiscal Year: 2013
    ..In addition, these studies could lead to strategies for the treatment or prevention of cancers caused by oncogenic viruses. Therefore, there are cogent and health-related justifications for these studies. ..
  21. Rediscovery of the Nucleolinus
    MARK COLLIN ALLIEGRO; Fiscal Year: 2012
    ..Such a finding would represent a fundamental advance in our understanding of centrosome biogenesis and have broad implications for cell and biomedical sciences. ..
  22. MICROTUBULE DYNAMICS AND MITOTIC MECHANISM
    Timothy J Mitchison; Fiscal Year: 2013
    ..We propose to test and extend this model. If confirmed, it will provide significant conceptual progress on early embryo organization. ..
  23. Comparative molecular physiology of mammalian formins
    Henry N Higgs; Fiscal Year: 2012
    ..Our findings provide new and exciting opportunities for therapies against pathologies involving malfunction of these mechanisms. ..
  24. Cell Free System for Identification of MHC Class II Immunodominant Epitopes
    Scheherazade Sadegh-Nasseri; Fiscal Year: 2011
    ..Thus, the new technology proposed in this project takes a major leap forward in predicting accurately immunodominant peptides. Project ..
  25. Mechanism of the eukaryotic replicative DNA helicase
    Johannes Walter; Fiscal Year: 2013
    ..Together, our experiments will elucidate the molecular mechanism and dynamics of the MCM2-7 helicase and thereby greatly deepen our knowledge of how cells copy and maintain their genomes. ) ..
  26. MATRIX BASED MINERAL ENAMEL-BIOMIMETICS
    JANET M OLDAK; Fiscal Year: 2013
    ..Moreover, this study will have a significant impact on the field of biomineralization, macromolecular self-assembly, protein structure, and the understanding of pathological enamel formation. ..
  27. REGULATION OF OXYGEN METABOLISM IN BLOOD NEUTROPHILS
    LINDA MCPHAIL; Fiscal Year: 2005
    ..Similarly, we will identify the oxidase component(s) that bind(s) DG and assess the role of DG-binding in NADPH oxidase activation. These studies should provide new insights into the regulation of NADPH oxidase activation. ..
  28. How does the KSHV small capsid protein function to promote self-assembly?
    Prashant Desai; Fiscal Year: 2013
    ..Specific Aim 2. Use a cell-free system and in vitro methods to discover the novel functions of the SCP. I. Cell-free assembly. II. Sedimentation analysis of sub-assemblies. III. In vitro MCP-scaffold protein binding assay. ..
  29. VIRAL PROTEINS IN ROTAVIRUS REPLICATION
    John Patton; Fiscal Year: 1991
    ..These results should provide valuable information on events in the replication of the rotaviruses that may be useful for developing strategies of controlling diseases caused by these viruses...
  30. REGULATION OF SUPEROXIDE PRODUCTION BY HUMAN NEUTROPHILS
    Jeffrey Friedman; Fiscal Year: 2007
    ..These studies may ultimately lead to new pharmacological methods for controlling oxygen radical-mediated tissue damage as well as an improved understanding of the pathophysiology of CGD. ..
  31. Molecular Mechanisms of ApoE4 Proteolysis in Alzheimer's Disease
    Troy T Rohn; Fiscal Year: 2013
    ..Both immunohistochemical analysis using fixed post mortem brain sections as well as Western blot analysis from AD samples will be performed and compared to age-matched controls. ..
  32. REGULATION OF GLCNAC-PI DEACETYLATION
    Victoria Stevens; Fiscal Year: 1999
    ..Finally, the protein that mediates the GTP effect will be identified and isolated by reconstituting this stimulation in microsomes in which it had been inactivated. ..
  33. Cell-free VEE assembly system and alphavirus drug screen
    JAISRI LINGAPPA; Fiscal Year: 2006
    ..The cell-free VEE assembly system offer a safe and low cost approach to understanding the mechanism of VEE capsid formation and to screen for agents active against alphaviruses that have potential as bioterrorism agents. ..
  34. PROTEOLYSIS IN HUMAN ERYTHROID CELLS
    JOSEPH SHAEFFER; Fiscal Year: 1991
    ..The proteolysis of excess, newly synthesized non-Alpha chains in the blood cells of individuals with Alpha-thalassemic disorders, e.g. HbH disease, will also be studied by these procedures...
  35. MEMBRANE FUSION ATPASES AND THE GOLGI APPARATUS
    Ayano Satoh; Fiscal Year: 2007
    ..Insights into the molecular mechanism of Golgi reassembly may therefore provide insight into these medical conditions. ..
  36. Role of Adapter Protein in Infectious Diseases
    Maria Diakonova; Fiscal Year: 2005
    ..abstract_text> ..
  37. FATTY ACID AND PEPTIDE AMIDATION--A SHARED MECHANSIM
    Gregory Mueller; Fiscal Year: 2003
    ..Demonstrating that oleamide biosynthesis proceeds through PAM, or via an alternative pathway, will provide a basis for improving the diagnosis and treatment of sleep disorders. ..
  38. MECHANISM OF AMINO ACID NEUROTRANSMITTER RELEASE
    Tetsufumi Ueda; Fiscal Year: 1993
    ....
  39. A cell-free system for exocytosis in the beta-cell
    SUSANNE STRAUB SHARP; Fiscal Year: 2005
    ..physiological mechanisms such as time-dependent potentiation and biphasic insulin release. The method will also allow the study of some aspects of endocytosis. ..
  40. METABOLIC ACTIVATION OF UNSUBSTITUTED HYDROXAMIC ACID
    MEI SIE LEE; Fiscal Year: 1990
    ..Results obtained from these studies will aid in the elucidation of activation mechanisms of this class of compounds...
  41. REGULATION OF NEUTROPHIL FUNCTION
    J Lambeth; Fiscal Year: 2000
    ..Techniques in use include lipid biochemistry, recombinant DNA methods, expression cloning, protein expression and protein purification. ..
  42. PROLIFERATIVE SIGNALS FROM IL-3 RECEPTORS TO NUCLEUS
    Theodore Gabig; Fiscal Year: 1993
    ..The long term goals of this study are to purify the most relevant or novel proteins involved in this signalling process, to clone and sequence their corresponding cDNA's, and define their function in the signalling process...
  43. MOLECULAR BASIS FOR MUSCLE PROTEIN LOSS IN CACHEXIA
    Stewart Lecker; Fiscal Year: 2003
    ..abstract_text> ..
  44. Mechanism of PCNA-Dependent Cdt1 Destruction in S phase
    COURTNEY HAVENS; Fiscal Year: 2009
    ..I propose to study in molecular detail how this destruction process takes place. Because Cdt1 over expression can cause cancer, the work is highly relevant for human health. ..
  45. ELUCIDATION OF THE MECHANISMS--IN VITRO GOLGI TRANSPORT
    MARGARET WEIDMAN; Fiscal Year: 1999
    ..A clearer understanding of the mechanisms and regulation of protein trafficking within the Golgi will contribute to the rational design of pharmacological therapeutics for these types of genetic diseases. ..
  46. TAILORING ANTICANCER THERAPY TO LOSS OF P53
    David Fisher; Fiscal Year: 2003
    ..abstract_text> ..
  47. A cell-free system for assaying ion channel function
    BENJAMIN DORANZ; Fiscal Year: 2003
    ..We have incorporated ion channels into lipoparticles, but have had no way of measuring their ability to function. We propose here to develop a nano-scale sensor for detecting ion channel function. ..
  48. IN VIVO CHEMILUMINESCENT ACTIVATION OF PHOTOSENSITIZERS
    Susan Carpenter; Fiscal Year: 1999
    ..reaction of firefly luciferase and luciferin can activate the antiviral activity of hypericin in a cell free system; however, the efficiency of energy transfer was found to depend on the localized concentrations of the energy ..
  49. CORONAVIRUS--ANALYSIS OF POLYMERASE GENE PRODUCTS
    Mark Denison; Fiscal Year: 1992
    ..Initial studies will involve translation of genomic RNA in vitro in a cell free system. This system will be used to analyze the size, kinetics, processing of gene A translation products and will ..
  50. SYNTHESIS AND DISTRIBUTION OF PROTEINS IN MEMBRANES
    David Sabatini; Fiscal Year: 2004
    ..For these studies we use a cell free system that we developed that recreates in vitro the generation of post-Golgi vesicles from a purified Golgi ..
  51. HOMOLOGOUS RECOMBINATION IN HUMAN CELL EXTRACTS
    Raju Kucherlapati; Fiscal Year: 1991
    ..The cDNA will be inserted into mammalian expression vectors and its ability to enhance homologous recombination will be studies. Isolation of a genomic clone corresponding to the cDNA and its characterization are also proposed...
  52. A cell-free system for assaying ion channel function
    BENJAMIN DORANZ; Fiscal Year: 2007
    ..In previous studies we demonstrated that ion channels could be incorporated into Lipoparticles, and in Phase 1 studies we demonstrated that ion channels could function within the Lipoparticle. ..
  53. Novel apoptotic pathway activated by misfolded proteins
    Dale Bredesen; Fiscal Year: 2007
    ..abstract_text> ..
  54. ACTIVATION AND REGULATION OF NORMAL AND NEOPLASTIC CELLS
    Stanley Cohen; Fiscal Year: 1992
    ..It is hoped that these studies will provide insights into basic mechanisms of cell growth and regulation...
  55. END JOINING REACTION IN DNA REPAIR & V(D)J RECOMBINATION
    Gilbert Chu; Fiscal Year: 2005
    ..Experiments will determine if DNA polymerase(s) are involved. The cell free system will be used to characterize the joining of hairpin ends, which occurs during V(D)J recombination, identify ..
  56. NEURONAL CELL DEATH
    RANDALL PITTMAN; Fiscal Year: 2004
    ..A cell free system has been developed in which cellular extracts from apoptotic and control cells can be treated with various ..
  57. REGULATION OF HEPATOMA ENZYME SYNTHESIS BY CYCLIC AMP
    WESLEY WICKS; Fiscal Year: 1980
    ..TAT inducibility by cAMP analogs and insulin will be examined in metaphase-arrested and enucleated H35 cells. Dex is known to be ineffective in both situations...
  58. BIOCHEM DISSECTION OF P53-DEPENDENT APOPTOSIS PATHWAY
    Han Fei Ding; Fiscal Year: 2003
    ..Dr. David E. Fisher will supervise this project and provide guidance in my transition to an independent investigator. ..
  59. BIOCHEMICAL ANALYSIS OF NUCLEAR STRUCTURE
    John Newport; Fiscal Year: 1999
    ..Successful completion of this work will provide a foundation for understanding how foci form and what role this subnuclear compartment plays in ensuring that DNA replication occurs successfully. ..
  60. ASSEMBLY OF HIV CAPSIDE IN A CELL FREE SYSTEM
    JAISRI LINGAPPA; Fiscal Year: 2001
    ..Aim #3 is to purify the TCP-1 related protein in order to obtain high affinity antibodies and functional proteins that can be used in inactivation, depletion and reconstitution experiments that define the TCP-1 role. ..
  61. INITIATION OF DNA REPLICATION BY PHOSPHORYLATION
    Martine Coue; Fiscal Year: 2005
    ..Thus, the results of the proposed experiments will shed light on the process of cell division, as it occurs normally during development and in healthy animals, and abnormally in disease processes such as cancer. ..
  62. Role of Mastermind in Notch Transcriptional Activation
    Joseph White; Fiscal Year: 2006
    ....
  63. CHARACTERIZATION GAMMA-SEC ACTIVITY CELL FREE SYSTEM
    David Hoke; Fiscal Year: 2004
    ..abstract_text> ..
  64. HBF VARIANTS FOR GENE THERAPY OF SICKLE CELL DISEASE
    Kazuhiko Adachi; Fiscal Year: 2005
    ..Using a rabbit reticulocyte cell free system, we will measure degradation of non-alpha chain in the absence of a chain during translation in the presence ..