directed molecular evolution

Summary

Summary: The techniques used to produce molecules exhibiting properties that conform to the demands of the experimenter. These techniques combine methods of generating structural changes with methods of selection. They are also used to examine proposed mechanisms of evolution under in vitro selection conditions.

Top Publications

  1. ncbi Artificial evolution of an enzyme active site: structural studies of three highly active mutants of Escherichia coli alkaline phosphatase
    M H Le Du
    Departement d Ingenierie et d Etudes des Proteines, CEA, Saclay, Gif sur Yvette, France
    J Mol Biol 316:941-53. 2002
  2. ncbi Design and evolution of new catalytic activity with an existing protein scaffold
    Hee-Sung Park
    Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 373-1, Kusung-dong, Yusung-gu, Daejon 305-701, Korea
    Science 311:535-8. 2006
  3. ncbi In the light of directed evolution: pathways of adaptive protein evolution
    Jesse D Bloom
    Division of Biology, California Institute of Technology, Pasadena, CA 91125, USA
    Proc Natl Acad Sci U S A 106:9995-10000. 2009
  4. ncbi Directed evolution of a monomeric, bright and photostable version of Clavularia cyan fluorescent protein: structural characterization and applications in fluorescence imaging
    Hui-wang Ai
    Department of Chemistry, University of Alberta, Edmonton, AB, Canada T6G 2G2
    Biochem J 400:531-40. 2006
  5. ncbi In-vitro protein evolution by ribosome display and mRNA display
    Dasa Lipovsek
    Biological Engineering Division, Massachusetts Institute of Technology, Cambridge 02139, USA
    J Immunol Methods 290:51-67. 2004
  6. ncbi Regulatory and metabolic rewiring during laboratory evolution of ethanol tolerance in E. coli
    Hani Goodarzi
    Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA
    Mol Syst Biol 6:378. 2010
  7. ncbi Directed molecular evolution in plant improvement
    M Lassner
    Maxygen Incorporated, 515 Galveston Drive, California 94063, Redwood City, USA
    Curr Opin Plant Biol 4:152-6. 2001
  8. ncbi Evolution of programmable zinc finger-recombinases with activity in human cells
    Russell M Gordley
    Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA
    J Mol Biol 367:802-13. 2007
  9. ncbi Genome shuffling leads to rapid phenotypic improvement in bacteria
    Ying-Xin Zhang
    Maxygen, 515 Galveston Drive, Redwood City, California 94063, USA
    Nature 415:644-6. 2002
  10. ncbi Discovery and directed evolution of a glyphosate tolerance gene
    Linda A Castle
    Verdia, Inc Redwood City, CA 94063, USA
    Science 304:1151-4. 2004

Research Grants

  1. Vaccines for Eastern and Western Equine Encephalitis Viruses
    Robert Whalen; Fiscal Year: 2007
  2. MECHANISTIC STUDIES OF PROKARYOTIC MANNOSYLTRANSFERASES
    JON THORSON; Fiscal Year: 2002
  3. STRUCTURE FUNCTION RELATIONSHIP OF TUMOR SUPPRESSORS
    Ming Daw Tsai; Fiscal Year: 1999
  4. Creation of Stable Cleaved Trimers of the HIV-1 Envelope Protein
    Robert Whalen; Fiscal Year: 2006
  5. Engineering high affinity integrin binding proteins
    Jennifer Cochran; Fiscal Year: 2007
  6. Dissecting Electrostatic Effects in Cystic Fibrosis Muco
    Gerard Wong; Fiscal Year: 2005
  7. A Novel Technology for Molecular Evolution
    JAMES LARRICK; Fiscal Year: 2003
  8. Engineering AAV for Enhanced Retrograde Transport
    Brian Kaspar; Fiscal Year: 2006
  9. Directed Evolution of Nucleic Acid Enzymes
    GERALD JOYCE; Fiscal Year: 2007
  10. Directed Evolution of Nucleic Acid Enzymes
    Gerald F Joyce; Fiscal Year: 2010

Detail Information

Publications188 found, 100 shown here

  1. ncbi Artificial evolution of an enzyme active site: structural studies of three highly active mutants of Escherichia coli alkaline phosphatase
    M H Le Du
    Departement d Ingenierie et d Etudes des Proteines, CEA, Saclay, Gif sur Yvette, France
    J Mol Biol 316:941-53. 2002
    ..This study shows how structural analysis may help to progress in the improvement of an enzyme catalytic activity (k(cat)), and explains the structural events associated with this artificial evolution...
  2. ncbi Design and evolution of new catalytic activity with an existing protein scaffold
    Hee-Sung Park
    Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 373-1, Kusung-dong, Yusung-gu, Daejon 305-701, Korea
    Science 311:535-8. 2006
    ..8 x 10(2) (mole/liter)(-1) second(-1), thus increasing resistance to Escherichia coli growth on cefotaxime by a factor of about 100...
  3. ncbi In the light of directed evolution: pathways of adaptive protein evolution
    Jesse D Bloom
    Division of Biology, California Institute of Technology, Pasadena, CA 91125, USA
    Proc Natl Acad Sci U S A 106:9995-10000. 2009
    ..These lessons about the coupling between adaptive and neutral protein evolution in the laboratory offer insight into the evolution of proteins in nature...
  4. ncbi Directed evolution of a monomeric, bright and photostable version of Clavularia cyan fluorescent protein: structural characterization and applications in fluorescence imaging
    Hui-wang Ai
    Department of Chemistry, University of Alberta, Edmonton, AB, Canada T6G 2G2
    Biochem J 400:531-40. 2006
    ..85) makes it particularly suitable as a replacement for ECFP (enhanced CFP) or Cerulean as a FRET (fluorescence resonance energy transfer) donor to either a yellow or orange FP acceptor...
  5. ncbi In-vitro protein evolution by ribosome display and mRNA display
    Dasa Lipovsek
    Biological Engineering Division, Massachusetts Institute of Technology, Cambridge 02139, USA
    J Immunol Methods 290:51-67. 2004
    ....
  6. ncbi Regulatory and metabolic rewiring during laboratory evolution of ethanol tolerance in E. coli
    Hani Goodarzi
    Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA
    Mol Syst Biol 6:378. 2010
    ..Remarkably, these laboratory-evolved strains, by and large, follow the same adaptive paths as inferred from our coarse-grained search of the fitness landscape...
  7. ncbi Directed molecular evolution in plant improvement
    M Lassner
    Maxygen Incorporated, 515 Galveston Drive, California 94063, Redwood City, USA
    Curr Opin Plant Biol 4:152-6. 2001
    b>Directed molecular evolution is a powerful tool to evolve genes with commercial applications...
  8. ncbi Evolution of programmable zinc finger-recombinases with activity in human cells
    Russell M Gordley
    Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA
    J Mol Biol 367:802-13. 2007
    ....
  9. ncbi Genome shuffling leads to rapid phenotypic improvement in bacteria
    Ying-Xin Zhang
    Maxygen, 515 Galveston Drive, Redwood City, California 94063, USA
    Nature 415:644-6. 2002
    ..This approach has the potential to facilitate cell and metabolic engineering and provide a non-recombinant alternative to the rapid production of improved organisms...
  10. ncbi Discovery and directed evolution of a glyphosate tolerance gene
    Linda A Castle
    Verdia, Inc Redwood City, CA 94063, USA
    Science 304:1151-4. 2004
    ..Glyphosate acetylation provides an alternative strategy for supporting glyphosate use on crops...
  11. ncbi HIV-1 proviral DNA excision using an evolved recombinase
    Indrani Sarkar
    Max Planck Institute for Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, D 01307 Dresden, Germany
    Science 316:1912-5. 2007
    ..Although a long way from use in the clinic, we speculate that this type of technology might be adapted in future antiretroviral therapies, among other possible uses...
  12. ncbi Rapid protein-folding assay using green fluorescent protein
    G S Waldo
    Structural Biology Group, MS M888, Los Alamos National Laboratory, NM 87545, USA
    Nat Biotechnol 17:691-5. 1999
    ..This approach to improving protein folding does not require functional assays for the protein of interest and provides a simple route to improving protein folding and expression by directed evolution...
  13. ncbi Directed evolution of high-affinity antibody mimics using mRNA display
    LiHui Xu
    Phylos, Inc, Lexington, MA 02421, USA
    Chem Biol 9:933-42. 2002
    ..10Fn3-based scaffold libraries and mRNA-display allow the isolation of high-affinity, specific antigen binding proteins; potential applications of such binding proteins include diagnostic protein microarrays and protein therapeutics...
  14. ncbi Optimization of human immunodeficiency virus type 1 envelope glycoproteins with V1/V2 deleted, using virus evolution
    Ilja Bontjer
    Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam CINIMA, Academic Medical Center, University of Amsterdam, Meibergdreef 15 K3 105, 1105 AZ Amsterdam, The Netherlands
    J Virol 83:368-83. 2009
    ..In general, virus evolution may provide a powerful tool to optimize Env vaccine antigens...
  15. ncbi Kemp elimination catalysts by computational enzyme design
    Daniela Röthlisberger
    Department of Biochemistry, University of Washington, Seattle, Washington 98195, USA
    Nature 453:190-5. 2008
    ..These results demonstrate the power of combining computational protein design with directed evolution for creating new enzymes, and we anticipate the creation of a wide range of useful new catalysts in the future...
  16. ncbi Compartmentalized self-replication: a novel method for the directed evolution of polymerases and other enzymes
    Farid J Ghadessy
    MRC Laboratory of Molecular Biology, Cambridge, UK
    Methods Mol Biol 352:237-48. 2007
    ..aquaticus Pol I (Taq) polymerase with novel and useful properties, such as increased thermostability or resistance to the potent inhibitor, heparin, from a repertoire of randomly mutated Taq polymerase genes...
  17. ncbi An enhanced system for unnatural amino acid mutagenesis in E. coli
    Travis S Young
    Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA
    J Mol Biol 395:361-74. 2010
    ..The versatility, increased yields, and increased stability of the pEVOL vector will further facilitate the expression of proteins with unnatural amino acids...
  18. ncbi A highly sensitive selection method for directed evolution of homing endonucleases
    Zhilei Chen
    Center for Biophysics and Computational Biology, University of Illinois, Urbana, IL 61801, USA
    Nucleic Acids Res 33:e154. 2005
    ..003%. This system should also be readily applicable for directed evolution of other DNA cleavage enzymes...
  19. ncbi Genome-wide detection of polymorphisms at nucleotide resolution with a single DNA microarray
    David Gresham
    Lewis Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ 08544, USA
    Science 311:1932-6. 2006
    ..We applied this approach to elucidate the genetic basis of phenotypic variants and to identify the small number of single-base pair changes accumulated during experimental evolution of yeast...
  20. ncbi Hsp90 as a capacitor for morphological evolution
    S L Rutherford
    Howard Hughes Medical Institute, University of Chicago, Illinois 60637, USA
    Nature 396:336-42. 1998
    ..This provides a plausible mechanism for promoting evolutionary change in otherwise entrenched developmental processes...
  21. ncbi Directed molecular evolution improves the immunogenicity and protective efficacy of a Venezuelan equine encephalitis virus DNA vaccine
    Lesley C Dupuy
    Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD 21702, USA
    Vaccine 27:4152-60. 2009
    We employed directed molecular evolution to improve the cross-reactivity and immunogenicity of the Venezuelan equine encephalitis virus (VEEV) envelope glycoproteins...
  22. ncbi Directed evolution of ATP binding proteins from a zinc finger domain by using mRNA display
    Glen S Cho
    Howard Hughes Medical Institute, Department of Molecular Biology, Massachusetts General Hospital, Boston, 02114, USA
    Chem Biol 13:139-47. 2006
    ..Many novel independent sequences were recovered with moderate affinity and high specificity for ATP, validating this scaffold for the generation of functional molecules...
  23. ncbi Evolution of variants of yeast site-specific recombinase Flp that utilize native genomic sequences as recombination target sites
    Swetha Bolusani
    School of Biological Sciences/IfM, 911 Hergot Avenue, Louisiana Tech University, Ruston, LA 71272, USA
    Nucleic Acids Res 34:5259-69. 2006
    ..We demonstrate the ability of an Flp variant to mediate integration of a reporter cassette in Escherichia coli via recombination at one of the IL10-derived sites...
  24. ncbi Iterative saturation mutagenesis (ISM) for rapid directed evolution of functional enzymes
    Manfred T Reetz
    Max Planck Institut fur Kohlenforschung, Kaiser Wilhelm Platz 1, 45470 Mulheim Ruhr, Germany
    Nat Protoc 2:891-903. 2007
    ..The pronounced increase in thermostability of the lipase from Bacillus subtilis (Lip A) as a result of applying ISM is illustrated here...
  25. ncbi In vitro evolution of a fungal laccase in high concentrations of organic cosolvents
    Miren Zumárraga
    Department of Biocatalysis, Institute of Catalysis, Consejo Superior de Investigaciones Cientificas, Cantoblanco, 28049 Madrid, Spain
    Chem Biol 14:1052-64. 2007
    ..Some mutations at the protein surface stabilized the laccase by allowing additional electrostatic and hydrogen bonding to occur...
  26. ncbi Test of synergistic interactions among deleterious mutations in bacteria
    S F Elena
    Center for Microbial Ecology, Michigan State University, East Lansing 48824, USA
    Nature 390:395-8. 1997
    ..Several pairs exhibit significant interactions for fitness, but they are antagonistic as often as they are synergistic. These results do not support the mutational deterministic hypothesis for the evolution of sex...
  27. ncbi Miniaturising the laboratory in emulsion droplets
    Andrew D Griffiths
    Institut de Science et d Ingénierie Supramoléculaires ISIS, 8 allee Gaspard Monge, BP 70028, F 67083 Strasbourg cedex, France
    Trends Biotechnol 24:395-402. 2006
    ..This review describes the scope and potential of IVC in areas such as in vitro evolution of proteins and RNAs, cell-free cloning and sequencing, genetics, genomics, and proteomics...
  28. ncbi High-throughput screening of enzyme libraries: in vitro evolution of a beta-galactosidase by fluorescence-activated sorting of double emulsions
    Enrico Mastrobattista
    Medical Research Council, Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, United Kingdom
    Chem Biol 12:1291-300. 2005
    ..Only two specific mutations were ever seen to provide this improvement in Ebg beta-galactosidase activity in vivo. In contrast, nearly all the improved beta-galactosidases selected in vitro resulted from different mutations...
  29. ncbi Genetic screens and directed evolution for protein solubility
    Geoffrey S Waldo
    BN-2, MS M888, Los Alamos National Laboratories, Los Alamos, NM 87545, USA
    Curr Opin Chem Biol 7:33-8. 2003
    ..Efficient, well-established strategies for generating and recombining genetic diversity, driven by new screening and selection methods, can furnish correctly folded, soluble protein...
  30. ncbi Diverse evolutionary trajectories characterize a community of RNA-cleaving deoxyribozymes: a case study into the population dynamics of in vitro selection
    Kenny Schlosser
    Department of Biochemistry and Biomedical Sciences and Department of Chemistry, McMaster University, Hamilton, Canada L8N 3Z5
    J Mol Evol 61:192-206. 2005
    ..This is the first study which thoroughly documents the topography of a deoxyribozyme fitness landscape over many generations of in vitro selection...
  31. ncbi Improving catalytic function by ProSAR-driven enzyme evolution
    Richard J Fox
    Codexis, Inc, 200 Penobscot Drive, Redwood City, California 94063, USA
    Nat Biotechnol 25:338-44. 2007
    ....
  32. ncbi Modifying the stereochemistry of an enzyme-catalyzed reaction by directed evolution
    Gavin J Williams
    School of Biochemistry and Molecular Biology, Department of Chemistry, University of Leeds, Leeds LS2 9JT, United Kingdom
    Proc Natl Acad Sci U S A 100:3143-8. 2003
    ..This demonstration is of considerable significance to synthetic chemists requiring efficient syntheses of complex stereoisomeric products, such as carbohydrate mimetics...
  33. ncbi Comparative genome sequencing of Escherichia coli allows observation of bacterial evolution on a laboratory timescale
    Christopher D Herring
    Department of Bioengineering, University of California, San Diego, California 92093, USA
    Nat Genet 38:1406-12. 2006
    ..The success of this new genome-scale approach indicates that real-time evolution studies will now be practical in a wide variety of contexts...
  34. ncbi Directed evolution of proteins for heterologous expression and stability
    Cintia Roodveldt
    Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel
    Curr Opin Struct Biol 15:50-6. 2005
    ....
  35. ncbi Site-saturation mutagenesis is more efficient than DNA shuffling for the directed evolution of beta-fucosidase from beta-galactosidase
    Monal R Parikh
    Department of Biochemistry, Center for Fundamental and Molecular Evolution, Emory University School of Medicine, Rollins Research Center, Room 4119, 1510 Clifton Road, Atlanta, GA 30322, USA
    J Mol Biol 352:621-8. 2005
    ..Site-saturation mutagenesis thus proved faster, less resource-intensive and more effective than DNA shuffling for this particular evolutionary pathway...
  36. ncbi Chemical and biochemical strategies for the randomization of protein encoding DNA sequences: library construction methods for directed evolution
    Cameron Neylon
    School of Chemistry, University of Southampton, Highfield SO17 1BJ, UK
    Nucleic Acids Res 32:1448-59. 2004
    b>Directed molecular evolution and combinatorial methodologies are playing an increasingly important role in the field of protein engineering...
  37. ncbi Nucleotide exchange and excision technology (NExT) DNA shuffling: a robust method for DNA fragmentation and directed evolution
    Kristian M Müller
    Institut fur Biologie III, Universität Freiburg Schänzlestrasse 1, 79104 Freiburg, Germany
    Nucleic Acids Res 33:e117. 2005
    ..1%). NExT DNA fragmentation is rational, easily executed and reproducible, making it superior to other techniques. Additionally, NExT could feasibly be applied to several other nucleotide analogs...
  38. ncbi Unbiased in vitro selection reveals the unique character of the self-cleaving antigenomic HDV RNA sequence
    Atef Nehdi
    RNA Group/Groupe ARN, , , , Sherbrooke, , Canada J1H 5N4
    Nucleic Acids Res 34:584-92. 2006
    ..Thus, just because a self-cleaving RNA motif is small does not imply that it occurs easily...
  39. ncbi In vitro selection of restriction endonucleases by in vitro compartmentalization
    Nobuhide Doi
    Department of Biosciences and Informatics, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan
    Nucleic Acids Res 32:e95. 2004
    ....
  40. ncbi Directed evolution by in vitro compartmentalization
    Oliver J Miller
    MRC Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 2QH, UK
    Nat Methods 3:561-70. 2006
  41. ncbi Narrowing substrate specificity in a directly evolved enzyme: the A293D mutant of aspartate aminotransferase
    Margaret A Chow
    Department of Molecular and Cell Biology, University of California, Berkeley, California 94720 3206, USA
    Biochemistry 43:12780-7. 2004
    ..While HEX is always in the closed conformation, HEX + A293D is observed in both the closed and a novel open conformation, allowing for more rapid product release...
  42. ncbi A ribozyme for the aldol reaction
    Stefan Fusz
    , , Gerhard-Domagk-Strasse 1, 53121 Bonn, Germany
    Chem Biol 12:941-50. 2005
    ....
  43. ncbi In vitro selection of small RNA-cleaving deoxyribozymes that cleave pyrimidine-pyrimidine junctions
    Kenny Schlosser
    Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Ontario, Canada
    Nucleic Acids Res 36:4768-77. 2008
    ..The same deoxyribozymes exhibited approximately 1000-fold lower activity against all RNA substrates, but could potentially be improved through further in vitro evolution and engineering...
  44. ncbi Protein engineering by cell-surface display
    K D Wittrup
    Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
    Curr Opin Biotechnol 12:395-9. 2001
    ..The promise of cell-surface display for directed evolution is being realized, with significant improvements recently reported in protein ligand binding affinity, stability, expression and enzymatic activity...
  45. ncbi RNA-catalyzed RNA polymerization: accurate and general RNA-templated primer extension
    W K Johnston
    Whitehead Institute for Biomedical Research, and Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02142, USA
    Science 292:1319-25. 2001
    ..Its polymerization is also quite accurate: when primers extended by 11 nucleotides were cloned and sequenced, 1088 of 1100 sequenced nucleotides matched the template...
  46. ncbi Optimization and optimality of a short ribozyme ligase that joins non-Watson-Crick base pairings
    M P Robertson
    Department of Chemistry and Biochemistry, Institute for Cellular and Molecular Biology, University of Texas at Austin, 78712, USA
    RNA 7:513-23. 2001
    ..The optimized L1 ligases may be optimal within their sequence spaces, and minimal ligases that span less than 60 nt in length have been engineered based on these results...
  47. ncbi Evolution of high mutation rates in experimental populations of E. coli
    P D Sniegowski
    Department of Biology, University of Pennsylvania, Philadelphia 19104, USA
    Nature 387:703-5. 1997
    ..Our results corroborate computer simulations of mutator evolution in adapting clonal populations, and may help to explain observations that associate high mutation rates with emerging pathogens and with certain cancers...
  48. ncbi Directed evolution of an enantioselective epoxide hydrolase: uncovering the source of enantioselectivity at each evolutionary stage
    Manfred T Reetz
    Max Planck Institut fur Kohlenforschung, Kaiser Wilhelm Platz 1, D 45470 Mulheim Ruhr, Germany
    J Am Chem Soc 131:7334-43. 2009
    ..Predictions regarding substrate scope and enantioselectivity of the best mutant are shown to be possible...
  49. ncbi A covalent chemical genotype-phenotype linkage for in vitro protein evolution
    Viktor Stein
    Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, CB2 1GA, Cambridge, UK
    Chembiochem 8:2191-4. 2007
  50. ncbi Alteration of Cre recombinase site specificity by substrate-linked protein evolution
    F Buchholz
    Hooper Research Foundation, University of California San Francisco UCSF, 513 Parnassus Ave, San Francisco, CA 94143 0552, USA
    Nat Biotechnol 19:1047-52. 2001
    b>Directed molecular evolution was applied to generate Cre recombinase variants that recognize a new DNA target sequence. Cre was adapted in a three-stage strategy to evolve recombinases to specifically recombine the new site...
  51. ncbi Tailoring in vitro evolution for protein affinity or stability
    L Jermutus
    Biochemisches Institut, , Winterthurerstrasse 190, , Switzerland
    Proc Natl Acad Sci U S A 98:75-80. 2001
    ..Therefore, by a combination of randomization and appropriate selection strategies, an in vitro evolution of protein properties in a predictable direction is possible...
  52. ncbi Forty years of in vitro evolution
    Gerald F Joyce
    Department of Chemistry and Molecular Biology, La Jolla, CA 92037, USA
    Angew Chem Int Ed Engl 46:6420-36. 2007
    ..This Review summarizes the concepts and methods for the directed evolution of RNA molecules in vitro...
  53. ncbi Exceptional convergent evolution in a virus
    J J Bull
    Department of Zoology, Institute of Cellular and Molecular Biology, University of Texas, Austin 78712, USA
    Genetics 147:1497-507. 1997
    ..Substitution rates and fitness improvements were higher during the initial period of adaptation than during a later period, except when the host was changed...
  54. ncbi DNA shuffling of a family of genes from diverse species accelerates directed evolution
    A Crameri
    Maxygen Inc, Santa Clara, California 95051, USA
    Nature 391:288-91. 1998
    ..Molecular breeding by shuffling can efficiently mix sequences from different species, unlike traditional breeding techniques. The power of family shuffling may arise from sparse sampling of a larger portion of sequence space...
  55. ncbi The effect of cytidine on the structure and function of an RNA ligase ribozyme
    J Rogers
    Department of Chemistry, The Scripps Research Institute, La Jolla, California 92037, USA
    RNA 7:395-404. 2001
    ....
  56. ncbi Directed evolution of restriction endonuclease BstYI to achieve increased substrate specificity
    James C Samuelson
    New England Biolabs, 32 Tozer Road, Beverly, MA 01915, USA
    J Mol Biol 319:673-83. 2002
    ..Moreover, cleavage of the GGATCC sequence is no longer detected. This study provides further evidence that laboratory evolution strategies offer a powerful alternative to structure-guided protein design...
  57. ncbi Directed evolution of RuBisCO hypermorphs through genetic selection in engineered E.coli
    Monal R Parikh
    Department of Biochemistry, Center for Fundamental and Applied Molecular Evolution, Emory University School of Medicine, Rollins Research Center, Room 4119, Atlanta, GA 30322, USA
    Protein Eng Des Sel 19:113-9. 2006
    ..These results demonstrate a new strategy for the artificial selection of RuBisCO and other non-native metabolic enzymes...
  58. ncbi A trans acting ribozyme that phosphorylates exogenous RNA
    Dayal Saran
    Department of Chemistry, Indiana University, 800 East Kirkwood Avenue, Bloomington, Indiana 47405, USA
    Biochemistry 44:15007-16. 2005
    ..Lack of reactivity in [Co(NH(3))(6)](3+) indicates a requirement for inner sphere contact with the metal ion, either for structural stabilization, catalysis, or both...
  59. ncbi Directed evolution of an amine oxidase for the preparative deracemisation of cyclic secondary amines
    Reuben Carr
    School of Chemistry, The University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JJ, Scotland, UK
    Chembiochem 6:637-9. 2005
  60. ncbi Altering protein specificity: techniques and applications
    Nina M Antikainen
    Department of Chemistry and Biochemistry, The Institute of Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX 78712, USA
    Bioorg Med Chem 13:2701-16. 2005
    ....
  61. ncbi General method for sequence-independent site-directed chimeragenesis
    Kaori Hiraga
    Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125, USA
    J Mol Biol 330:287-96. 2003
    ..These results illustrate the use of SISDC in creating designed chimeric protein libraries and further illustrate the ability of SCHEMA to identify chimeras whose folded structures are likely not to be disrupted by recombination...
  62. ncbi Expanding the genetic code
    Lei Wang
    The Jack H. Skirball Center for Chemical Biology and Proteomics, The Salk Institute for Biological Studies, La Jolla, California 92037, USA
    Annu Rev Biophys Biomol Struct 35:225-49. 2006
    ..This methodology provides a powerful tool both for exploring protein structure and function in vitro and in vivo and for generating proteins with new or enhanced properties...
  63. ncbi Advances in laboratory evolution of enzymes
    Shimon Bershtein
    Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel
    Curr Opin Chem Biol 12:151-8. 2008
    ....
  64. ncbi Development of a novel continuous culture device for experimental evolution of bacterial populations
    E de Crecy
    Evolugate 5745 SW 75th St 188, Gainesville, FL 32608, USA
    Appl Microbiol Biotechnol 77:489-96. 2007
    ..In 200 generations over 2 weeks of dilution cycles, the evolved strain improved in generation time by a factor of three, with no contaminations and easy manipulation...
  65. ncbi Functional analysis of organophosphorus hydrolase variants with high degradation activity towards organophosphate pesticides
    Catherine Mee-Hie Cho
    Department of Chemical and Environmental Engineering, University of California, Riverside, 92521, USA
    Protein Eng Des Sel 19:99-105. 2006
    ..Using structural modeling, these two mutations were shown to favor the formation of hydrogen bonds with nearby residues, resulting in structural changes that could alter the overall substrate hydrolysis...
  66. ncbi Evolutional design of a hyperactive cysteine- and methionine-free mutant of Escherichia coli dihydrofolate reductase
    Masahiro Iwakura
    National Institute of Advanced Industrial Science and Technology, 1 1 1 Higashi, Tsukuba, Ibaraki 305 8566, Japan
    J Biol Chem 281:13234-46. 2006
    ....
  67. ncbi Increased folding stability of TEM-1 beta-lactamase by in vitro selection
    Insa Kather
    Laboratorium für Biochemie und Bayreuther, Zentrum für Molekulare Biowissenschaften, Universitat Bayreuth, 95440 Bayreuth, Germany
    J Mol Biol 383:238-51. 2008
    ..Such unfavorable van der Waals repulsions are not easily identified in crystal structures or by computational approaches, but they strongly reduce the conformational stability of a protein...
  68. ncbi Modulation of effector affinity by hinge region mutations also modulates switching activity in an engineered allosteric TEM1 beta-lactamase switch
    Jin Ryoun Kim
    Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 3400 N. Charles Street, Baltimore, MD 21218, USA
    Arch Biochem Biophys 446:44-51. 2006
    ....
  69. ncbi High-throughput screening of enzyme libraries: thiolactonases evolved by fluorescence-activated sorting of single cells in emulsion compartments
    Amir Aharoni
    Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel
    Chem Biol 12:1281-9. 2005
    ....
  70. ncbi Frame shuffling: a novel method for in vitro protein evolution
    Kenji Kashiwagi
    Department of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, Ariake, Koto-ku, Tokyo, Japan
    Protein Eng Des Sel 19:135-40. 2006
    ..The resultant progeny produce mutant proteins having segmental sequence changes. Such frame-shuffled mutant proteins exhibit physicochemical properties that differ from those of proteins obtained using conventional mutagenesis...
  71. ncbi Loop grafting of Bacillus subtilis lipase A: inversion of enantioselectivity
    Ykelien L Boersma
    Department of Pharmaceutical Biology, Groningen University Institute for Drug Exploration GUIDE, University of Groningen, A Deusinglaan 1, 9713 AV Groningen, The Netherlands
    Chem Biol 15:782-9. 2008
    ..9% to +6.0%, whereas the cutinase-derived variant was improved to an ee of +26.5%. The enantioselectivity of the cutinase-derived variant was further improved by directed evolution to an ee of +57.4%...
  72. ncbi Combinatorial library approaches for improving soluble protein expression in Escherichia coli
    Darren J Hart
    EMBL Grenoble Outstation, 6 rue Jules Horowitz, 38042 Grenoble, France
    Acta Crystallogr D Biol Crystallogr 62:19-26. 2006
    ..This article reviews the progress in this field and provides a general overview of relevant mutation methods, screens and selections...
  73. ncbi Directed evolution of a maltogenic alpha-amylase from Bacillus sp. TS-25
    Aubrey Jones
    Novozymes Inc, 1445 Drew Avenue, Davis, CA 95616, USA
    J Biotechnol 134:325-33. 2008
    ..Relative to wild-type Novamyl, a number of the resulting variants exhibited more than 10 degrees C increase in thermal stability at pH 4.5, one of which demonstrated substantial anti-staling properties in low pH breads...
  74. ncbi Directed evolution of a beta-galactosidase from Pyrococcus woesei resulting in increased thermostable beta-glucuronidase activity
    Ai Sheng Xiong
    Biotechnology Research Institute, Shanghai Academy of Agricultural Sciences, 2901 Beidi Road, Shanghai, 201106, China
    Appl Microbiol Biotechnol 77:569-78. 2007
    ....
  75. ncbi Multivariate-activity mining for molecular quasi-species in a glutathione transferase mutant library
    Sanela Kurtovic
    Department of Biochemistry and Organic Chemistry, Uppsala University, BMC, Box 576, SE 75123 Uppsala, Sweden
    Protein Eng Des Sel 20:243-56. 2007
    ..The data show that multivariate analysis of functional profiles can identify small structural changes influencing the evolution of enzymes with novel substrate-activity profiles...
  76. ncbi Structural determinants of glutathione transferases with azathioprine activity identified by DNA shuffling of alpha class members
    Sanela Kurtovic
    Department of Biochemistry and Organic Chemistry, Uppsala University, BMC, Box 576, SE 75123 Uppsala, Sweden
    J Mol Biol 375:1365-79. 2008
    ..Knowledge of activity-determining segments in the structure is valuable in the protein engineering of glutathione transferase for enhanced or suppressed activity...
  77. ncbi Directed evolution of a non-heme-iron-dependent extradiol catechol dioxygenase: identification of mutants with intradiol oxidative cleavage activity
    Janne Schlosrich
    Department of Chemistry, University of Warwick, Coventry, CV4 7AL, UK
    Chembiochem 7:1899-908. 2006
    ....
  78. ncbi Directed evolution of aldolases for exploitation in synthetic organic chemistry
    Amanda Bolt
    School of Chemistry, University of Leeds, Leeds LS2 9JT, UK
    Arch Biochem Biophys 474:318-30. 2008
    ..The review also describes some of the fundamental insights into mechanistic enzymology that directed evolution can provide...
  79. ncbi Modified substrate specificity of pyrroloquinoline quinone glucose dehydrogenase by biased mutation assembling with optimized amino acid substitution
    Norio Hamamatsu
    Novartis Pharma K.K, Tsukuba Research Institute, Ohkubo 8, Tsukuba 300-2611, Japan
    Appl Microbiol Biotechnol 73:607-17. 2006
    ....
  80. ncbi Enhancement of the activity and alkaline pH stability of Thermobifida fusca xylanase A by directed evolution
    Qin Wang
    College of Life Science and Biotechnology, Shanghai Jiaotong University, 800 Rm 211, No 3 Biopharmacology Building, 800 Dongchuan Road, Shanghai, 200240, P R China
    Biotechnol Lett 30:937-44. 2008
    ..This study shows a useful approach to improve the catalytic activity and alkaline pH stability of T. fusca xylanase A toward the hydrolysis of xylan...
  81. ncbi Natural history as a predictor of protein evolvability
    Wayne M Patrick
    Department of Biochemistry, Center for Fundamental and Applied Molecular Evolution, Emory University, Atlanta, GA 30322, USA
    Protein Eng Des Sel 19:439-42. 2006
    ..Three examples of 'generalist' proteins that evolved in the laboratory into 'specialists' are described to illustrate the practical utility of this point...
  82. ncbi Improvement of the activity of arylmalonate decarboxylase by random mutagenesis
    Y Terao
    Department of Biosciences and Informatics, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan
    Appl Microbiol Biotechnol 73:647-53. 2006
    ..The activity of the triple mutant enzyme was tenfold higher than that of the starting doubly mutated enzyme...
  83. ncbi Protein engineering of hydrogenase 3 to enhance hydrogen production
    Toshinari Maeda
    Artie McFerrin Department of Chemical Engineering, Texas A and M University, College Station, TX 77843 3122, USA
    Appl Microbiol Biotechnol 79:77-86. 2008
    ..This is the first random protein engineering of a hydrogenase...
  84. ncbi Enhancing thermostability of Escherichia coli phytase AppA2 by error-prone PCR
    Moon Soo Kim
    Department of Animal Science and Graduate Field of Food Science, Cornell University, Ithaca, NY 14853, USA
    Appl Microbiol Biotechnol 79:69-75. 2008
    ..5, respectively. Thus, the catalytic efficiency of these enzymes was not inversely related to their thermostability...
  85. ncbi Creation of novel enantioselective lipases by SIMPLEX
    Yuichi Koga
    Department of Material and Life Science, Graduate School of Engineering, Osaka University, Osaka, Japan
    Methods Mol Biol 375:165-81. 2007
    ..Here, we describe the detail procedure to construct such an exclusively in vitro protein library and a practical screening method based on enzymatic activity...
  86. ncbi Improving evolution
    Nicole Rusk
    Nat Methods 3:242. 2006
    ..By combining elements of protein engineering and directed evolution, researchers open the door to creating enzymes with diverse catalytic functions in a protein scaffold of their choice...
  87. ncbi Directed evolution of transketolase substrate specificity towards an aliphatic aldehyde
    Edward G Hibbert
    Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, UK
    J Biotechnol 134:240-5. 2008
    ..This suggests that saturation mutagenesis can be more selectively guided for evolution towards either natural or non-natural substrates, using both structural and sequence information...
  88. ncbi Directed evolution of beta-galactosidase from Escherichia coli into beta-glucuronidase
    Ai Sheng Xiong
    College of Horticulture, Nanjing Agricultural University, Nanjing, China
    J Biochem Mol Biol 40:419-25. 2007
    ..The comparison of molecular models of the mutated and wildtype enzymes revealed the relationship between protein function and structural modification...
  89. ncbi Milestones in directed enzyme evolution
    Haiyan Tao
    Department of Chemistry, Columbia University, New York, NY 10027, USA
    Curr Opin Chem Biol 6:858-64. 2002
    ....
  90. ncbi Directed evolution of a bacterial alpha-amylase: toward enhanced pH-performance and higher specific activity
    Cornelius Bessler
    Henkel KGaA, Henkelstrasse 67, , Germany
    Protein Sci 12:2141-9. 2003
    ..In addition, three further mutations were found K406R, N414S, and E356D, the latter being present in other bacterial alpha-amylases...
  91. ncbi Demonstration of the importance and usefulness of manipulating non-active-site residues in protein design
    A Shimotohno
    Department of Biology, Graduate School of Science, Osaka University, Toyonaka, Osaka 560 0043, Japan
    J Biochem 129:943-8. 2001
    ..The present results illustrate how a protein function is dramatically modified by the accumulation of many seemingly inert mutations of non-active-site residues...
  92. ncbi Directed evolution of enzymes for applied biocatalysis
    Nicholas J Turner
    School of Chemistry, University of Edinburgh, King s Buildings, West Mains Road, Edinburgh EH9 3JJ, Scotland, UK
    Trends Biotechnol 21:474-8. 2003
    ....
  93. ncbi Laboratory evolution of cytochrome p450 BM-3 monooxygenase for organic cosolvents
    Tuck Seng Wong
    Division of Chemistry and Chemical Engineering 210-41, California Institute of Technology, Pasadena, California 91125, USA
    Biotechnol Bioeng 85:351-8. 2004
    ..The engineered p450 BM-3's are also significantly more resistant to acetone, acetonitrile, dimethylformamide, and ethanol as cosolvents in the reaction...
  94. ncbi Different strategies to recover the activity of monomeric triosephosphate isomerase by directed evolution
    G Saab-Rincon
    Instituto de Biotecnologia, UNAM, Apartado Postal 510 3, Cuernavaca, Morelos, 62271, Mexico
    Protein Eng 14:149-55. 2001
    ..A small difference in growth rate is observed when both mutant genes are compared, which seems to be attributable to a difference in solubility of the expressed proteins...
  95. ncbi Directed evolution relieves product inhibition and confers in vivo function to a rationally designed tyrosine aminotransferase
    Steven C Rothman
    Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720 3206, USA
    Protein Sci 13:763-72. 2004
    ..Both residues are in close proximity to Arg292 and the mutations may function to modulate the arginine switch mechanism responsible for dual substrate recognition in TATases and HEX...
  96. ncbi In vitro evolution of beta-glucuronidase into a beta-galactosidase proceeds through non-specific intermediates
    I Matsumura
    Institute of Cellular and Molecular Biology, ICMB A4800 MBB 3 424, University of Texas, 26th and Speedway, Austin, TX, 78712, USA
    J Mol Biol 305:331-9. 2001
    ..These results are consistent with the "patchwork" hypothesis, which postulates that modern enzymes diverged from ancestors with broad specificity...
  97. ncbi Combinatorial engineering to enhance amylosucrase performance: construction, selection, and screening of variant libraries for increased activity
    Bart A van der Veen
    , UMR CNRS 5504, UMR INRA 792, INSA, 135 Avenue de Rangueil, 31077 Toulouse Cedex 4, France
    FEBS Lett 560:91-7. 2004
    ..This allows discrimination between hydrolysis and transglucosylation, enabling a more detailed comparison between wild-type and mutant enzymes...
  98. ncbi Novel concepts for selection of catalytic activity
    P Soumillion
    , Institut des Sciences de la Vie, , 1 Place Louis Pasteur, B 1348 Louvain la-Neuve, Belgium
    Curr Opin Biotechnol 12:387-94. 2001
    ..The results achieved demonstrate the enormous potential of the methods and leave questions open for further studies...
  99. ncbi Stepwise manipulation of DNA specificity in Flp recombinase: progressively adapting Flp to individual and combinatorial mutations in its target site
    Yuri Voziyanov
    Section of Molecular Genetics and Microbiology, University of Texas, Austin, TX 78712 1095, USA
    J Mol Biol 326:65-76. 2003
    ..Our results suggest that combined DNA shuffling and mutagenesis of libraries of Flp variants active on distinct mFRTs can yield variants that can recombine mFRTs containing combinations of the individual mutations...
  100. ncbi In vitro selection of fibronectin gain-of-function mutations
    Patricia H Tani
    Department of Biochemistry and Molecular Biology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73190, USA
    Biochem J 365:287-94. 2002
    ..Additionally, these higher-affinity 3fn9-10 ligands provide a starting point for further in vitro evolution and engineering of integrin-specific modules...
  101. ncbi Dispelling the myths--biocatalysis in industrial synthesis
    Hans E Schoemaker
    DSM Research, Life Science Products, Post Office Box 18, 6160 MD Geleen, Netherlands
    Science 299:1694-7. 2003
    ..As a result, we expect many new industrial applications of biocatalysis to be realized, from single-step enzymatic conversions to customized multistep microbial synthesis by means of metabolic pathway engineering...

Research Grants96

  1. Vaccines for Eastern and Western Equine Encephalitis Viruses
    Robert Whalen; Fiscal Year: 2007
    ..glycoproteins of VEEV have already been obtained using in vitro multigene DNA recombination and directed molecular evolution. This demonstrates the potential of this approach for improving EEEV and WEEV immunogens...
  2. MECHANISTIC STUDIES OF PROKARYOTIC MANNOSYLTRANSFERASES
    JON THORSON; Fiscal Year: 2002
    ....
  3. STRUCTURE FUNCTION RELATIONSHIP OF TUMOR SUPPRESSORS
    Ming Daw Tsai; Fiscal Year: 1999
    ..The goal is to provide a structural model for the suppressor cdk4 complex, which will be useful in deciphering the "missing functionality" in the disease-related mutants of tumor suppressors, an important information for drug design. ..
  4. Creation of Stable Cleaved Trimers of the HIV-1 Envelope Protein
    Robert Whalen; Fiscal Year: 2006
    ..We propose to use directed molecular evolution to accomplish 2 main objectives...
  5. Engineering high affinity integrin binding proteins
    Jennifer Cochran; Fiscal Year: 2007
    ..period of the award, where integrin binding proteins will be engineered to even higher affinity using directed molecular evolution by yeast surface display...
  6. Dissecting Electrostatic Effects in Cystic Fibrosis Muco
    Gerard Wong; Fiscal Year: 2005
    ..3) To develop, via directed molecular evolution, an anionic DNAzyme which mimics the biofilm suppression action of cationic lactoferrin, but will not ..
  7. A Novel Technology for Molecular Evolution
    JAMES LARRICK; Fiscal Year: 2003
    ..These will be expressed, purified and characterized by binding and affinity measurements. This simple yet high-yield system should contribute significantly to proteomics research in the post-genome era. ..
  8. Engineering AAV for Enhanced Retrograde Transport
    Brian Kaspar; Fiscal Year: 2006
    ..We therefore propose to employ a directed molecular evolution approach to develop novel AAV capsid variants with enhanced retrograde transport in vivo...
  9. Directed Evolution of Nucleic Acid Enzymes
    GERALD JOYCE; Fiscal Year: 2007
    ..These decorated nanostructures then will be subject to in vitro evolution, selecting on the basis of both their form and the function of the appended DNAs. ..
  10. Directed Evolution of Nucleic Acid Enzymes
    Gerald F Joyce; Fiscal Year: 2010
    ..These decorated nanostructures then will be subject to in vitro evolution, selecting on the basis of both their form and the function of the appended DNAs. ..
  11. Directed Evolution of Nucleic Acid Enzymes
    GERALD JOYCE; Fiscal Year: 2005
    ..The adapter will allow the cofactor to be bound readily by the enzyme, allowing evolution to exploit the bound cofactor for use in catalysis. ..
  12. Synthetic Protein Families by Structure-Guided SCHEMA Recombination
    Frances Arnold; Fiscal Year: 2009
    ..Finally, this research represents a fundamentally new approach to study protein stability using recombination, with results that can be applied to understanding enzymes in general. ..
  13. The Combinatorial Design of Protein Molecular Switches
    Marc Ostermeier; Fiscal Year: 2009
    ..We will develop switches designed for the treatment of cancer and switches designed to be used as sensors for the detection and quantification of important protein kinases in vitro and in live cells. ..
  14. Live Cell Fluorescence Imaging Using Molecular Switches
    Marc Ostermeier; Fiscal Year: 2007
    ....
  15. The Combinatorial Design of Protein Molecular Switches
    Marc Ostermeier; Fiscal Year: 2007
    ..abstract_text> ..
  16. The Combinatorial Design of Protein Molecular Switches
    MARC A OSTERMEIER; Fiscal Year: 2010
    ..We will develop switches designed for the treatment of cancer and switches designed to be used as sensors for the detection and quantification of important protein kinases in vitro and in live cells. ..
  17. Synthetic Protein Families by Structure-Guided SCHEMA Recombination
    Frances H Arnold; Fiscal Year: 2010
    ..Finally, this research represents a fundamentally new approach to study protein stability using recombination, with results that can be applied to understanding enzymes in general. ..
  18. Creation of site-specific DNA methyltransferases
    Marc Ostermeier; Fiscal Year: 2006
    ..abstract_text> ..
  19. Quantitative Microfluidic Molecular Evolution
    BRIAN PAEGEL; Fiscal Year: 2006
    ....
  20. Computation-Guided Protein Recombination and Evolution
    Frances Arnold; Fiscal Year: 2006
    ..abstract_text> ..
  21. Mammalian Genomes - Stasis and Change
    HOLLY WICHMAN; Fiscal Year: 2009
    ..This proposal will also ask whether changes in patterns of methylation by endocrine disruptors is linked to increased rates of retrotransposition. ..
  22. Ribozymes for Peptide- and Protein- Sensing Chip Arrays
    Andrew Ellington; Fiscal Year: 2004
    ..1.-D.3), and (2)Adapting peptide and protein-activated aptazymes to function in chip arrays (D.4). ..
  23. LEDGF-Integrase Structural Biology
    Alan Engelman; Fiscal Year: 2005
    ..abstract_text> ..
  24. Interrogation of systems level mechanisms controlling DNA repair processes
    Nitin S Baliga; Fiscal Year: 2010
    ..e. a predictive model for regulatory mechanisms for repair. This basic model will serve as a template for designing systems approaches to model higher complexities of eukaryotic repair processes. ..
  25. MicroRNA Profiles of Pathogen Infection
    Andrew Ellington; Fiscal Year: 2005
    ..3. Apply these methods to the detection of changes in microRNA expression during influenza infection. 4. Apply these methods to the detection of changes in microRNA expression with other infectious agents. ..
  26. REV DECOYS FOR GENE THERAPY AND DRUG DEVELOPMENT
    Andrew Ellington; Fiscal Year: 1999
    ..In particular, the peptide motifs identified by physical mapping and selection studies should provide information essential to the synthesis of peptidomimetic drugs. ..
  27. Interrogation of systems level mechanisms controlling DNA repair processes
    NITIN BALIGA; Fiscal Year: 2007
    ..e. a predictive model for regulatory mechanisms for repair. This basic model will serve as a template for designing systems approaches to model higher complexities of eukaryotic repair processes. ..
  28. REV DECOYS FOR GENE THERAPY AND DRUG DEVELOPMENT
    Andrew Ellington; Fiscal Year: 2004
    ....
  29. Site-specific incorporation of FRET pairs into intracellular proteins
    Andrew Ellington; Fiscal Year: 2009
    ..This will allow us to follow and track proteins in a cell, and to determine where, when, and how proteins reside next to one another, in either normal or malformed complexes. ..