Juan S Bonifacino

Summary

Affiliation: National Institutes of Health
Country: USA

Publications

  1. pmc Role of the mammalian retromer in sorting of the cation-independent mannose 6-phosphate receptor
    Cecilia N Arighi
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    J Cell Biol 165:123-33. 2004
  2. pmc Multilayered mechanism of CD4 downregulation by HIV-1 Vpu involving distinct ER retention and ERAD targeting steps
    Javier G Magadán
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, United States of America
    PLoS Pathog 6:e1000869. 2010
  3. pmc Human Vam6p promotes lysosome clustering and fusion in vivo
    S Caplan
    Cell Biology and Metabolism Branch at the National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    J Cell Biol 154:109-22. 2001
  4. pmc Recognition of dileucine-based sorting signals from HIV-1 Nef and LIMP-II by the AP-1 gamma-sigma1 and AP-3 delta-sigma3 hemicomplexes
    Katy Janvier
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    J Cell Biol 163:1281-90. 2003
  5. pmc Stonin 2: an adaptor-like protein that interacts with components of the endocytic machinery
    J A Martina
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    J Cell Biol 153:1111-20. 2001
  6. pmc GGAs: a family of ADP ribosylation factor-binding proteins related to adaptors and associated with the Golgi complex
    E C Dell'Angelica
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    J Cell Biol 149:81-94. 2000
  7. ncbi request reprint Ubiquitin and the control of protein fate in the secretory and endocytic pathways
    J S Bonifacino
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892 5430, USA
    Annu Rev Cell Dev Biol 14:19-57. 1998
  8. ncbi request reprint Insights into the biogenesis of lysosome-related organelles from the study of the Hermansky-Pudlak syndrome
    Juan S Bonifacino
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, Building 18T Room 101, National Institutes of Health, Bethesda, MD 20892, USA
    Ann N Y Acad Sci 1038:103-14. 2004
  9. pmc Retromer
    Juan S Bonifacino
    Cell Biology and Metabolism Program, National Institute of Child Health and Human Development, Building 18T Room 101, National Institutes of Health, Bethesda, MD 20892, USA
    Curr Opin Cell Biol 20:427-36. 2008
  10. ncbi request reprint The GGA proteins: adaptors on the move
    Juan S Bonifacino
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, Building 18T Room 101, National Institutes of Health, Bethesda, Maryland 20892, USA
    Nat Rev Mol Cell Biol 5:23-32. 2004

Detail Information

Publications117 found, 100 shown here

  1. pmc Role of the mammalian retromer in sorting of the cation-independent mannose 6-phosphate receptor
    Cecilia N Arighi
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    J Cell Biol 165:123-33. 2004
    ..These observations indicate that retromer prevents the delivery of the CI-MPR to lysosomes, probably by sequestration into endosome-derived tubules from where the receptor returns to the TGN...
  2. pmc Multilayered mechanism of CD4 downregulation by HIV-1 Vpu involving distinct ER retention and ERAD targeting steps
    Javier G Magadán
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, United States of America
    PLoS Pathog 6:e1000869. 2010
    ..The multiple levels at which Vpu engages these cellular quality control mechanisms underscore the importance of ensuring profound suppression of CD4 to the life cycle of HIV-1...
  3. pmc Human Vam6p promotes lysosome clustering and fusion in vivo
    S Caplan
    Cell Biology and Metabolism Branch at the National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    J Cell Biol 154:109-22. 2001
    ..This study implicates hVam6p as a mammalian tethering/docking factor characterized with intrinsic ability to promote lysosome fusion in vivo...
  4. pmc Recognition of dileucine-based sorting signals from HIV-1 Nef and LIMP-II by the AP-1 gamma-sigma1 and AP-3 delta-sigma3 hemicomplexes
    Katy Janvier
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    J Cell Biol 163:1281-90. 2003
    ..These observations reveal a novel mode of recognition of sorting signals involving the gamma/delta and sigma subunits of AP-1 and AP-3...
  5. pmc Stonin 2: an adaptor-like protein that interacts with components of the endocytic machinery
    J A Martina
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    J Cell Biol 153:1111-20. 2001
    ..These observations suggest that stonin 2 is a novel component of the general endocytic machinery...
  6. pmc GGAs: a family of ADP ribosylation factor-binding proteins related to adaptors and associated with the Golgi complex
    E C Dell'Angelica
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    J Cell Biol 149:81-94. 2000
    ..Disruption of both GGA genes in yeast resulted in impaired trafficking of carboxypeptidase Y to the vacuole. These observations suggest that GGAs are components of ARF-regulated coats that mediate protein trafficking at the TGN...
  7. ncbi request reprint Ubiquitin and the control of protein fate in the secretory and endocytic pathways
    J S Bonifacino
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892 5430, USA
    Annu Rev Cell Dev Biol 14:19-57. 1998
    ..These recent findings imply that ubiquitin plays more diverse roles in the regulation of the fate of cellular proteins than originally anticipated...
  8. ncbi request reprint Insights into the biogenesis of lysosome-related organelles from the study of the Hermansky-Pudlak syndrome
    Juan S Bonifacino
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, Building 18T Room 101, National Institutes of Health, Bethesda, MD 20892, USA
    Ann N Y Acad Sci 1038:103-14. 2004
    ..Further studies of these complexes should contribute to the elucidation of the mechanisms of LRO biogenesis and the pathogenesis of HPS...
  9. pmc Retromer
    Juan S Bonifacino
    Cell Biology and Metabolism Program, National Institute of Child Health and Human Development, Building 18T Room 101, National Institutes of Health, Bethesda, MD 20892, USA
    Curr Opin Cell Biol 20:427-36. 2008
    ..Recent studies have implicated retromer in a broad range of physiological, developmental and pathological processes, underscoring the critical nature of retrograde transport mediated by this complex...
  10. ncbi request reprint The GGA proteins: adaptors on the move
    Juan S Bonifacino
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, Building 18T Room 101, National Institutes of Health, Bethesda, Maryland 20892, USA
    Nat Rev Mol Cell Biol 5:23-32. 2004
    ....
  11. ncbi request reprint The mechanisms of vesicle budding and fusion
    Juan S Bonifacino
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Cell 116:153-66. 2004
    ..Precise regulation of these two aspects of vesicular transport ensures efficient cargo transfer while preserving organelle identity...
  12. pmc A diacidic motif in human immunodeficiency virus type 1 Nef is a novel determinant of binding to AP-2
    O Wolf Lindwasser
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, Building 18T, Room 101, National Institutes of Health, Bethesda, MD 20892, USA
    J Virol 82:1166-74. 2008
    ....
  13. pmc Transport according to GARP: receiving retrograde cargo at the trans-Golgi network
    Juan S Bonifacino
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Trends Cell Biol 21:159-67. 2011
    ..Loss of GARP function compromises the growth, fertility and/or viability of the defective organisms, emphasizing the essential nature of GARP-mediated retrograde transport...
  14. ncbi request reprint Signals for sorting of transmembrane proteins to endosomes and lysosomes
    Juan S Bonifacino
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA
    Annu Rev Biochem 72:395-447. 2003
    ..This complex array of signals and recognition proteins ensures the dynamic but accurate distribution of transmembrane proteins to different compartments of the endosomal-lysosomal system...
  15. ncbi request reprint Retrograde transport from endosomes to the trans-Golgi network
    Juan S Bonifacino
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA
    Nat Rev Mol Cell Biol 7:568-79. 2006
    ..We also propose a central role for a 'tubular endosomal network' in sorting to recycling pathways that lead not only to the trans-Golgi network but also to different plasma-membrane domains and to specialized storage vesicles...
  16. ncbi request reprint CD4 down-regulation by HIV-1 and simian immunodeficiency virus (SIV) Nef proteins involves both internalization and intracellular retention mechanisms
    Jeremy J Rose
    Laboratory of Molecular Microbiology, NIAID, National Institutes of Health, Bethesda, Maryland 20892, USA
    J Biol Chem 280:7413-26. 2005
    ..In contrast, AP-2-mediated enhanced endocytosis constitutes the predominant mechanism for SIV (MAC-239 strain) Nef-induced down-regulation of human CD4 in human cells...
  17. ncbi request reprint The trihelical bundle subdomain of the GGA proteins interacts with multiple partners through overlapping but distinct sites
    Rafael Mattera
    Cell Biology and Metabolism Branch, NICHD, National Institutes of Health, Bethesda, Maryland 20892, USA
    J Biol Chem 279:31409-18. 2004
    ..The different GAT binding partners engage in both competitive and cooperative interactions that may be important for the function of the GGAs in protein sorting...
  18. ncbi request reprint Definition of the consensus motif recognized by gamma-adaptin ear domains
    Rafael Mattera
    Cell Biology and Metabolism Branch, NICHD, National Institutes of Health, Bethesda, Maryland 20892, USA
    J Biol Chem 279:8018-28. 2004
    ..These findings shed light on the mechanism of accessory protein recruitment to trans-Golgi network and endosomal clathrin coats...
  19. pmc Assembly of the biogenesis of lysosome-related organelles complex-3 (BLOC-3) and its interaction with Rab9
    Daniel P Kloer
    Laboratory of Molecular Biology, NIDDK, Eunice Kennedy Shriver NICHD, National Institutes of Health, Bethesda, Maryland 20892, USA
    J Biol Chem 285:7794-804. 2010
    ..This interaction is mediated by HPS4 and the switch I and II regions of Rab9. These characteristics indicate that BLOC-3 might function as a Rab9 effector in the biogenesis of LROs...
  20. pmc Downregulation of CD4 by human immunodeficiency virus type 1 Nef is dependent on clathrin and involves direct interaction of Nef with the AP2 clathrin adaptor
    Rittik Chaudhuri
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, Bldg 18T, Rm 101, National Institutes of Health, Bethesda, MD 20892, USA
    J Virol 81:3877-90. 2007
    ..Together, these results support a model in which HIV-1 Nef downregulates CD4 by promoting its accelerated endocytosis by a clathrin/AP2 pathway...
  21. pmc Coatomer-dependent protein delivery to lipid droplets
    Krishnakant G Soni
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    J Cell Sci 122:1834-41. 2009
    ..Here we demonstrate that ATGL and ADRP, but not TIP47, are delivered to LDs by a pathway mediated by the COPI and COPII coatomer proteins and their corresponding regulators...
  22. pmc Adaptor protein 2-mediated endocytosis of the β-secretase BACE1 is dispensable for amyloid precursor protein processing
    Yogikala Prabhu
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Mol Biol Cell 23:2339-51. 2012
    ..Taken together, these observations support the notion that BACE1 is capable of cleaving APP in late compartments of the secretory pathway...
  23. ncbi request reprint An ear-core interaction regulates the recruitment of the AP-3 complex to membranes
    Stephane Lefrancois
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Dev Cell 7:619-25. 2004
    ..These observations suggest a new regulatory mechanism for the recruitment of AP-3 to membranes involving delta-ear-sigma3 interactions...
  24. ncbi request reprint Recognition of accessory protein motifs by the gamma-adaptin ear domain of GGA3
    Gregory J Miller
    Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Department of Health and Human Services, Bethesda, Maryland 20892, USA
    Nat Struct Biol 10:599-606. 2003
    ....
  25. pmc Ang2/fat-free is a conserved subunit of the Golgi-associated retrograde protein complex
    F Javier Pérez-Victoria
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Mol Biol Cell 21:3386-95. 2010
    ..These findings indicate that Ang2 is the missing component of the GARP complex in most eukaryotes...
  26. pmc Requirement of the human GARP complex for mannose 6-phosphate-receptor-dependent sorting of cathepsin D to lysosomes
    F Javier Pérez-Victoria
    Cell Biology and Metabolism Program, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Mol Biol Cell 19:2350-62. 2008
    ..We also report that a Vps54 mutant protein in the Wobbler mouse strain is active in retrograde transport, thus explaining the viability of these mutant mice...
  27. pmc Structural mechanism for ubiquitinated-cargo recognition by the Golgi-localized, gamma-ear-containing, ADP-ribosylation-factor-binding proteins
    Gali Prag
    Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA
    Proc Natl Acad Sci U S A 102:2334-9. 2005
    ..This ability highlights the GAT domain as a hub for interactions with multiple partners in trafficking...
  28. pmc The trans-Golgi network accessory protein p56 promotes long-range movement of GGA/clathrin-containing transport carriers and lysosomal enzyme sorting
    Gonzalo A Mardones
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Mol Biol Cell 18:3486-501. 2007
    ..We conclude that p56 tightly cooperates with the GGAs in the sorting of cathepsin D to lysosomes, probably by enabling the movement of GGA-containing TCs...
  29. pmc Interchangeable but essential functions of SNX1 and SNX2 in the association of retromer with endosomes and the trafficking of mannose 6-phosphate receptors
    Raul Rojas
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, Building 18T Room 101, National Institutes of Health, Bethesda, MD 20892, USA
    Mol Cell Biol 27:1112-24. 2007
    ....
  30. pmc Structural basis for ubiquitin recognition and autoubiquitination by Rabex-5
    Sangho Lee
    Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, US National Institutes of Health NIH, Bethesda, Maryland 20892, USA
    Nat Struct Mol Biol 13:264-71. 2006
    ..The A20 zinc-finger diaromatic patch mediates ubiquitin-ligase activity by directly recruiting a ubiquitin-loaded ubiquitin-conjugating enzyme...
  31. pmc Morphology and dynamics of clathrin/GGA1-coated carriers budding from the trans-Golgi network
    Rosa Puertollano
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development and Laboratory of Cell Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA
    Mol Biol Cell 14:1545-57. 2003
    ....
  32. pmc A basic patch on alpha-adaptin is required for binding of human immunodeficiency virus type 1 Nef and cooperative assembly of a CD4-Nef-AP-2 complex
    Rittik Chaudhuri
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Building 18T, Room 101, National Institutes of Health, Bethesda, MD 20892, USA
    J Virol 83:2518-30. 2009
    ..This cooperativity explains how Nef is able to efficiently downregulate CD4 despite weak binary interactions between components of the tripartite complex...
  33. ncbi request reprint Interactions of GGA3 with the ubiquitin sorting machinery
    Rosa Puertollano
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA
    Nat Cell Biol 6:244-51. 2004
    ..The VHS and GAT domains of GGA3 are responsible for this binding, as well as for interactions with TSG101, a component of the ubiquitin-dependent sorting machinery. Thus, GGAs may have additional roles in sorting of ubiquitinated cargo...
  34. ncbi request reprint Phosphoregulation of sorting signal-VHS domain interactions by a direct electrostatic mechanism
    Yukio Kato
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA
    Nat Struct Biol 9:532-6. 2002
    ....
  35. pmc Canonical interaction of cyclin G associated kinase with adaptor protein 1 regulates lysosomal enzyme sorting
    Satoshi Kametaka
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, and Laboratory of Cell Biology, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA
    Mol Biol Cell 18:2991-3001. 2007
    ..Thus, interactions between the PsiG[PDE][PsiLM]-motif sequences in GAK and the AP1-gamma-ear domain are critical for the recruitment of GAK to the TGN and the function of GAK in lysosomal enzyme sorting...
  36. pmc The Vps27/Hse1 complex is a GAT domain-based scaffold for ubiquitin-dependent sorting
    Gali Prag
    Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, U S Department of Health and Human Services, Bethesda, MD 20892, USA
    Dev Cell 12:973-86. 2007
    ..Coarse-grained Monte Carlo simulations of the Vps27/Hse1 complex on a membrane show how the complex binds cooperatively to lipids and ubiquitinated membrane proteins and acts as a scaffold for ubiquitination reactions...
  37. pmc Sorting of the Alzheimer's disease amyloid precursor protein mediated by the AP-4 complex
    Patricia V Burgos
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Dev Cell 18:425-36. 2010
    ....
  38. pmc Regulation of retromer recruitment to endosomes by sequential action of Rab5 and Rab7
    Raul Rojas
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    J Cell Biol 183:513-26. 2008
    ..These findings demonstrate that Rab5 and Rab7 act in concert to regulate retromer recruitment to endosomes...
  39. ncbi request reprint BLOC-3, a protein complex containing the Hermansky-Pudlak syndrome gene products HPS1 and HPS4
    Jose A Martina
    Cell Biology and Metabolism Branch, NICHD, National Institutes of Health, Bethesda, Maryland 20892, USA
    J Biol Chem 278:29376-84. 2003
    ....
  40. pmc Functional architecture of the retromer cargo-recognition complex
    Aitor Hierro
    Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, US Department of Health and Human Services, Bethesda, Maryland 20892, USA
    Nature 449:1063-7. 2007
    ..This extended structure presents multiple binding sites for the SNX complex and receptor cargo, and appears capable of flexing to conform to curved vesicular membranes...
  41. pmc Gga2 mediates sequential ubiquitin-independent and ubiquitin-dependent steps in the trafficking of ARN1 from the trans-Golgi network to the vacuole
    Yi Deng
    Liver Diseases Branch, NIDDK, National Institutes of Health, Bethesda, Maryland 20892, USA
    J Biol Chem 284:23830-41. 2009
    ..These studies suggest that Gga2 is involved in sorting at both the TGN and multivesicular body and that the first step can occur without ubiquitin binding...
  42. ncbi request reprint Structural basis for acidic-cluster-dileucine sorting-signal recognition by VHS domains
    Saurav Misra
    Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA
    Nature 415:933-7. 2002
    ..The rigid spatial alignment of the three binding subsites leads to high specificity...
  43. ncbi request reprint The Rab5 guanine nucleotide exchange factor Rabex-5 binds ubiquitin (Ub) and functions as a Ub ligase through an atypical Ub-interacting motif and a zinc finger domain
    Rafael Mattera
    Cell Biology and Metabolism Branch, NICHD, National Institutes of Health, Bethesda, Maryland 20892
    J Biol Chem 281:6874-83. 2006
    ..Moreover, the demonstration that Rabex-5 is a ubiquitin ligase that binds ubiquitin and undergoes ubiquitination indicates that its role in endosome fusion may be subject to additional regulation by ubiquitin-dependent modifications...
  44. pmc Human immunodeficiency virus type 1 Nef protein targets CD4 to the multivesicular body pathway
    Luis L P daSilva
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Building 18T, Room 101, National Institutes of Health, Bethesda, MD 20892, USA
    J Virol 83:6578-90. 2009
    ..We also show that both CD4 and Nef are ubiquitinated on lysine residues, but this modification is dispensable for Nef-induced targeting of CD4 to the MVB pathway...
  45. pmc Adaptor and clathrin exchange at the plasma membrane and trans-Golgi network
    Xufeng Wu
    Laboratory of Cell Biology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA
    Mol Biol Cell 14:516-28. 2003
    ..We conclude that clathrin-coated pits are dynamic structures with rapid exchange of both clathrin and adaptors and that adaptors are able to exchange independently of clathrin when clathrin exchange is blocked...
  46. pmc Structural basis for the wobbler mouse neurodegenerative disorder caused by mutation in the Vps54 subunit of the GARP complex
    F Javier Pérez-Victoria
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Proc Natl Acad Sci U S A 107:12860-5. 2010
    ..Severely reduced levels of mutant Vps54 and, consequently, of the whole GARP complex underlie the phenotype of the wobbler mouse...
  47. pmc Involvement of clathrin and AP-2 in the trafficking of MHC class II molecules to antigen-processing compartments
    Peter J McCormick
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Proc Natl Acad Sci U S A 102:7910-5. 2005
    ..Because AP-2 is only associated with the plasma membrane, these results also indicate that a significant pool of MHC-II molecules traffic to the endosomal-lysosomal system by means of the cell surface...
  48. pmc Direct binding to Rsp5p regulates ubiquitination-independent vacuolar transport of Sna3p
    Hadiya Watson
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Mol Biol Cell 18:1781-9. 2007
    ..Our data demonstrate that Sna3p follows a novel ubiquitination-independent, but Rsp5p-mediated, sorting pathway to the vacuole...
  49. pmc A tubular EHD1-containing compartment involved in the recycling of major histocompatibility complex class I molecules to the plasma membrane
    Steve Caplan
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    EMBO J 21:2557-67. 2002
    ..These observations suggest an additional function of EHD1 as a tubule-inducing factor in the Arf6 pathway for recycling of plasma membrane proteins internalized by clathrin-independent endocytosis...
  50. ncbi request reprint Mechanisms of CD4 downregulation by the Nef and Vpu proteins of primate immunodeficiency viruses
    O Wolf Lindwasser
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Curr Mol Med 7:171-84. 2007
    ..These two mechanisms lead to a dramatic reduction of CD4 expression in infected cells and are essential for efficient virus replication and disease progression...
  51. pmc Ubiquitin binding and conjugation regulate the recruitment of Rabex-5 to early endosomes
    Rafael Mattera
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    EMBO J 27:2484-94. 2008
    ..We also show that monoubiquitinated Rabex-5 is enriched in the cytosol. These observations are consistent with a model whereby a cycle of Ub binding and monoubiquitination regulates the association of Rabex-5 with endosomes...
  52. ncbi request reprint Specific regulation of the adaptor protein complex AP-3 by the Arf GAP AGAP1
    Zhongzhen Nie
    Laboratory of Cellular Oncology, Center for Cancer Research, National Cancer Institute, Building 37, Room 4118, Bethesda, MD 20892, USA
    Dev Cell 5:513-21. 2003
    ..Cells overexpressing AGAP1 also exhibit increased LAMP1 trafficking via the plasma membrane. Taken together, these results support the hypothesis that AGAP1 directly and specifically regulates AP-3-dependent trafficking...
  53. pmc Divalent interaction of the GGAs with the Rabaptin-5-Rabex-5 complex
    Rafael Mattera
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    EMBO J 22:78-88. 2003
    ..These observations thus identify a binding sequence for GAE/gamma-adaptin ear domains and reveal a functional link between proteins regulating TGN cargo export and endosomal tethering/fusion events...
  54. pmc The adaptor protein-1 μ1B subunit expands the repertoire of basolateral sorting signal recognition in epithelial cells
    Xiaoli Guo
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Dev Cell 27:353-66. 2013
    ..We conclude that expression of distinct μ1 isoforms in epithelial cells expands the repertoire of signals recognized by AP-1 for sorting of a broader range of cargoes to the basolateral surface. ..
  55. pmc Role of the endocytic machinery in the sorting of lysosome-associated membrane proteins
    Katy Janvier
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Mol Biol Cell 16:4231-42. 2005
    ..Because AP-2 is exclusively associated with plasma membrane clathrin coats, our observations imply that a significant population of Lamps traffic via the plasma membrane en route to lysosomes...
  56. pmc The clathrin adaptor complexes as a paradigm for membrane-associated allostery
    Bertram J Canagarajah
    Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA
    Protein Sci 22:517-29. 2013
    ..The details of these mechanisms show how membrane docking and allosteric activation of AP complexes are intimately connected...
  57. pmc Transmembrane domain determinants of CD4 Downregulation by HIV-1 Vpu
    Javier G Magadán
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA
    J Virol 86:757-72. 2012
    ..These findings highlight the exploitation of several TMD-mediated mechanisms by HIV-1 Vpu in order to downregulate CD4 and thus promote viral pathogenesis...
  58. pmc Epidermal growth factor-dependent phosphorylation of the GGA3 adaptor protein regulates its recruitment to membranes
    Satoshi Kametaka
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, Building 18T Room 101, National Institutes of Health, Bethesda, MD 20892, USA
    Mol Cell Biol 25:7988-8000. 2005
    ..These observations indicate that EGF signaling elicits phosphorylation events that regulate the association of GGA3 with organellar membranes...
  59. pmc Conservation and diversification of dileucine signal recognition by adaptor protein (AP) complex variants
    Rafael Mattera
    Cell Biology and Metabolism Program, NICHD, National Institutes of Health, Bethesda, Maryland 20892, USA
    J Biol Chem 286:2022-30. 2011
    ..However, the characteristics of the binding site on each complex vary, providing for the specific recognition of a diverse repertoire of (D/E)XXXL(L/I) signals...
  60. ncbi request reprint Lysosome-related organelles
    E C Dell'Angelica
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892 5430, USA
    FASEB J 14:1265-78. 2000
    ....
  61. ncbi request reprint Coat proteins: shaping membrane transport
    Juan S Bonifacino
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, Building 18T Room 101, National Institutes of Health, Bethesda, Maryland 20892, USA
    Nat Rev Mol Cell Biol 4:409-14. 2003
    ....
  62. pmc Genomic screen for vacuolar protein sorting genes in Saccharomyces cerevisiae
    Cecilia J Bonangelino
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA
    Mol Biol Cell 13:2486-501. 2002
    ..The identification and phenotypic characterization of these novel mutants provide new insights into the mechanisms of vacuolar protein sorting, most notably the probable involvement of the actin cytoskeleton in this process...
  63. pmc Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1
    Xuefeng Ren
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Cell 152:755-67. 2013
    ..A third Arf1 interaction site near the N terminus of the γ subunit is important for recruitment, but not activation. These observations lead to a model for the recruitment and activation of AP-1 by Arf1...
  64. pmc Serine residues in the cytosolic tail of the T-cell antigen receptor alpha-chain mediate ubiquitination and endoplasmic reticulum-associated degradation of the unassembled protein
    Shuhei Ishikura
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver NICHD, Bethesda, MD 20892, USA
    J Biol Chem 285:23916-24. 2010
    ..We also found that this ubiquitination was mediated by the ER-localized ubiquitin ligase, HRD1. These findings indicate that serine-dependent, HRD1-mediated ubiquitination targets TCRalpha to the ERAD pathway...
  65. ncbi request reprint In vitro assays of Arf1 interaction with GGA proteins
    Hye Young Yoon
    Laboratory of Cellular Oncology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA
    Methods Enzymol 404:316-32. 2005
    ..In this chapter, we describe in vitro assays for analysis of GGA interaction with Arf1 x GTP and for determining intracellular Arf1 x GTP levels...
  66. pmc The retromer subunit Vps26 has an arrestin fold and binds Vps35 through its C-terminal domain
    Hang Shi
    Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, US National Institutes of Health, US Department of Health and Human Services, Bethesda, Maryland 20892, USA
    Nat Struct Mol Biol 13:540-8. 2006
    ..Hydrophobic residues and a glycine in this loop are required for integration into the retromer complex and endosomal localization of human Vps26, and for the function of yeast Vps26 in carboxypeptidase Y sorting...
  67. pmc Adaptins: the final recount
    M Boehm
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Mol Biol Cell 12:2907-20. 2001
    ..Finally, we extend our survey to adaptin-related proteins such as the GGAs and stonins, which contain domains homologous to the adaptins...
  68. pmc Crystallographic and functional analysis of the ESCRT-I /HIV-1 Gag PTAP interaction
    Young Jun Im
    Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892 0580, USA
    Structure 18:1536-47. 2010
    ..However, the mutant alleles did rescue downregulation of endogenous EGF receptor. This demonstrates that the PSAP motif is not rate determining in EGF receptor downregulation under normal conditions...
  69. pmc Structural requirements for function of yeast GGAs in vacuolar protein sorting, alpha-factor maturation, and interactions with clathrin
    C Mullins
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892 5430, USA
    Mol Cell Biol 21:7981-94. 2001
    ..Thus, these observations demonstrate that the yeast GGAs play important roles in the CPY pathway, vacuole biogenesis, and alpha-factor maturation and identify structural determinants that are critical for these functions...
  70. pmc Signal-mediated, AP-1/clathrin-dependent sorting of transmembrane receptors to the somatodendritic domain of hippocampal neurons
    Ginny G Farías
    Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Neuron 75:810-23. 2012
    ..These findings highlight the primary recognition event that underlies somatodendritic sorting and contribute to the evolving view of AP-1 as a global regulator of cell polarity...
  71. ncbi request reprint The contribution of VHL substrate binding and HIF1-alpha to the phenotype of VHL loss in renal cell carcinoma
    Jodi K Maranchie
    Urologic Oncology Branch, National Institutes of Health, Bethesda, MD 20892, USA
    Cancer Cell 1:247-55. 2002
    ..We further demonstrate that normoxic stabilization of HIF1alpha alone, while capable of mimicking some aspects of VHL loss, is not sufficient to reproduce tumorigenesis, indicating that it is not the critical oncogenic substrate of VHL...
  72. pmc Dual roles of the mammalian GARP complex in tethering and SNARE complex assembly at the trans-golgi network
    F Javier Pérez-Victoria
    Cell Biology and Metabolism Program, NICHD, Building 18T Room 101, NIH, Bethesda, MD 20892, USA
    Mol Cell Biol 29:5251-63. 2009
    ..These findings indicate that GARP orchestrates retrograde transport from endosomes to the TGN by promoting vesicle tethering and assembly of SNARE complexes in consecutive, independent steps...
  73. ncbi request reprint Pallidin is a component of a multi-protein complex involved in the biogenesis of lysosome-related organelles
    Kengo Moriyama
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Traffic 3:666-77. 2002
    ..These observations indicate that pallidin is a subunit of a novel multi-protein complex involved in the biogenesis of lysosome-related organelles...
  74. ncbi request reprint Distinct requirements for the AP-3 adaptor complex in pigment granule and synaptic vesicle biogenesis in Drosophila melanogaster
    C Mullins
    Cell Biology and Metabolism Branch, National Institutes of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Mol Gen Genet 263:1003-14. 2000
    ..Together, these results demonstrate that Drosophila AP-3 is critical for the biogenesis of pigment granules, but is apparently not essential for formation of a major population of synaptic vesicles in vivo...
  75. ncbi request reprint Defective expression of the mu3 subunit of the AP-3 adaptor complex in the Drosophila pigmentation mutant carmine
    C Mullins
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Mol Gen Genet 262:401-12. 1999
    ..These findings provide evidence of a role for mu3 in the sorting processes required for pigment granule biogenesis in Drosophila...
  76. ncbi request reprint Molecular characterization of hepatocystin, the protein that is defective in autosomal dominant polycystic liver disease
    Joost P H Drenth
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Building 18T, Room 101, Bethesda, MD 20892, USA
    Gastroenterology 126:1819-27. 2004
    ..Thus, the exact localization and cellular function of hepatocystin remain unclear...
  77. ncbi request reprint Genetic analyses of adaptin function from yeast to mammals
    Markus Boehm
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, Building 18T Room 101, National Institutes of Health, Bethesda, MD 20892, USA
    Gene 286:175-86. 2002
    ..Here, we review recent progress in the genetic analysis of the function of AP complexes, focusing on studies that make use of targeted interference or naturally-occurring mutations in various model organisms...
  78. ncbi request reprint Sorting of mannose 6-phosphate receptors mediated by the GGAs
    R Puertollano
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Science 292:1712-6. 2001
    ..A dominant-negative GGA mutant blocked exit of the receptors from the trans-Golgi network. Thus, the GGAs appear to mediate sorting of the mannose 6-phosphate receptors at the trans-Golgi network...
  79. ncbi request reprint The molecular machinery for lysosome biogenesis
    C Mullins
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Bioessays 23:333-43. 2001
    ..Other recently identified components, however, appear to be unique to higher eukaryotes. BioEssays 23:333-343, 2001. Published 2001 John Wiley & Sons, Inc...
  80. pmc Altered expression of a novel adaptin leads to defective pigment granule biogenesis in the Drosophila eye color mutant garnet
    C E Ooi
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    EMBO J 16:4508-18. 1997
    ..Thus, the eye pigmentation defect in the Drosophila garnet mutant may be attributed to compromised function of a coat protein involved in intracellular transport processes required for biogenesis or function of pigment granules...
  81. ncbi request reprint Altered trafficking of lysosomal proteins in Hermansky-Pudlak syndrome due to mutations in the beta 3A subunit of the AP-3 adaptor
    E C Dell'Angelica
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA
    Mol Cell 3:11-21. 1999
    ..Our results suggest that AP-3 functions in protein sorting to lysosomes and provide an example of a human disease in which altered trafficking of integral membrane proteins is due to mutations in a component of the sorting machinery...
  82. ncbi request reprint The GGAs promote ARF-dependent recruitment of clathrin to the TGN
    R Puertollano
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, Bethesda, MD 20892, USA
    Cell 105:93-102. 2001
    ..These observations suggest that the GGAs could function to link clathrin to membrane-bound ARF.GTP...
  83. pmc Functional and physical interactions of the adaptor protein complex AP-4 with ADP-ribosylation factors (ARFs)
    M Boehm
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    EMBO J 20:6265-76. 2001
    ..These observations establish AP-4 as an ARF1 effector and suggest a novel mode of interaction between ARF1 and an AP complex involving both constitutive and regulated interactions...
  84. ncbi request reprint Interaction of tyrosine-based sorting signals with clathrin-associated proteins
    H Ohno
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Science 269:1872-5. 1995
    ..The interaction was confirmed by in vitro binding assays. Thus, it is likely that the medium chains serve as signal-binding components of the clathrin-dependent sorting machinery...
  85. ncbi request reprint Cloning of the gene encoding the murine clathrin-associated adaptor medium chain mu 2: gene organization, alternative splicing and chromosomal assignment
    H Ohno
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Gene 210:187-93. 1998
    ..The isolation and characterization of the mu 2 gene should be instrumental for future studies of the genetics and physiological role of the adaptor medium chains in mammals...
  86. ncbi request reprint Signal-binding specificity of the mu4 subunit of the adaptor protein complex AP-4
    R C Aguilar
    Cell Biology and Metabolism Branch and the Laboratory of Molecular Genetics, NICHD, National Institutes of Health, Bethesda, Maryland 20892, USA
    J Biol Chem 276:13145-52. 2001
    ..This chimera was targeted to the endosomal-lysosomal system without being internalized from the plasma membrane...
  87. ncbi request reprint AP-4, a novel protein complex related to clathrin adaptors
    E C Dell'Angelica
    Cell Biology and Metabolism Branch, NICHD, National Institutes of Health, Bethesda, Maryland 20892, USA
    J Biol Chem 274:7278-85. 1999
    ..We propose that, like the related AP-1, AP-2, and AP-3 complexes, AP-4 plays a role in signal-mediated trafficking of integral membrane proteins in mammalian cells...
  88. ncbi request reprint Endosome-specific localization and function of the ARF activator GNOM
    Juan S Bonifacino
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Cell 112:141-2. 2003
    ..In this issue of Cell, Geldner et al. demonstrate that the Arabidopsis ARF activator GNOM localizes to endosomes where it controls the polarized trafficking of the auxin efflux carrier PIN1 to the basal plasma membrane...
  89. pmc ADP-Ribosylation factor 1 (ARF1) regulates recruitment of the AP-3 adaptor complex to membranes
    C E Ooi
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health NIH, Bethesda, Maryland 20892, USA
    J Cell Biol 142:391-402. 1998
    ..This finding suggests that ARF1 is not a regulator of specific coat proteins, but rather is a ubiquitous molecular switch that acts as a transducer of diverse signals influencing coat assembly...
  90. ncbi request reprint Immunoprecipitation
    J S Bonifacino
    National Institute of Child Health and Human Development, Bethesda, Maryland, USA
    Curr Protoc Cell Biol . 2001
    ..An additional method is provided for increasing the specificity of the technique by reprecipitating the antigen with the same or a different antibody...
  91. ncbi request reprint T cell CD3-zeta eta heterodimer expression and coupling to phosphoinositide hydrolysis
    M Mercep
    Biological Response Modifiers Program, National Cancer Institute, Bethesda, MD 20892
    Science 242:571-4. 1988
    ....
  92. pmc AP-3: an adaptor-like protein complex with ubiquitous expression
    E C Dell'Angelica
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    EMBO J 16:917-28. 1997
    ..These results suggest that the sigma3 chains are components of a novel, ubiquitous adaptor-like complex involved in the recognition of tyrosine-based sorting signals...
  93. ncbi request reprint Molecular characterization of the protein encoded by the Hermansky-Pudlak syndrome type 1 gene
    E C Dell'Angelica
    Cell Biology and Metabolism Branch, NICHD, National Institutes of Health, Bethesda, Maryland 20892, USA
    J Biol Chem 275:1300-6. 2000
    ....
  94. pmc Regulating the retention of T-cell receptor alpha chain variants within the endoplasmic reticulum: Ca(2+)-dependent association with BiP
    C K Suzuki
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892
    J Cell Biol 114:189-205. 1991
    ..These data suggest that the intracellular fate of newly synthesized proteins stably associated with BiP can be regulated by Ca2+ levels in the ER...
  95. ncbi request reprint Association of the AP-3 adaptor complex with clathrin
    E C Dell'Angelica
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Science 280:431-4. 1998
    ..AP-3 colocalized with clathrin in cells as observed by immunofluorescence and immunoelectron microscopy. Thus, AP-3 function in protein sorting may depend on clathrin...
  96. ncbi request reprint Metabolic labeling with amino acids
    J S Bonifacino
    National Institute of Child Health and Human Development, Bethesda, Maryland, USA
    Curr Protoc Mol Biol . 2001
    ....
  97. ncbi request reprint Interaction of endocytic signals from the HIV-1 envelope glycoprotein complex with members of the adaptor medium chain family
    H Ohno
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA
    Virology 238:305-15. 1997
    ..These observations suggest that HIV-1 Env utilizes the protein sorting machinery of the host cells for internalization and sorting at various steps of the endocytic and biosynthetic pathways...
  98. ncbi request reprint Beta3A-adaptin, a subunit of the adaptor-like complex AP-3
    E C Dell'Angelica
    Cell Biology and Metabolism Branch, NICHD, National Institutes of Health, Bethesda, Maryland 20892, USA
    J Biol Chem 272:15078-84. 1997
    ..The characteristics of beta3A-adaptin reported here lend support to the idea that AP-3 is a structural and functional homolog of the clathrin-associated adaptors AP-1 and AP-2...
  99. ncbi request reprint Localization of endogenous furin in cultured cell lines
    J Shapiro
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    J Histochem Cytochem 45:3-12. 1997
    ..Therefore, our results suggest that endogenous furin is predominantly localized to the area of the Golgi complex, most likely within the trans-Golgi network...
  100. ncbi request reprint Immunoprecipitation
    J S Bonifacino
    National Institute of Child Health and Human Development, Bethesda, Maryland, USA
    Curr Protoc Mol Biol . 2001
    ..g., with and without detergent, using glass beads, etc.). Flow charts and figures give the user a clear-cut explanation of the options for employing the technology...
  101. ncbi request reprint Biosynthetic labeling of proteins
    J S Bonifacino
    National Institute of Child Health and Human Development, Bethesda, Maryland, USA
    Curr Protoc Immunol . 2001
    ..Label incorporation can be determined as described in a support protocol. The cells used in these protocols can be either from suspension cultures or from single-cell suspensions of spleen or thymus...