Home LiteratureArticle Details
PMID: 9693377 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

ATP- and cytosol-dependent release of adaptor proteins from clathrin-coated vesicles: A dual role for Hsc70.

Molecular biology of the cell ·Vol. 9 ·No. 8 ·1998-08-00 ·Pages 2217-29

Hannan LA, Newmyer SL, Schmid SL

Abstract

Clathrin-coated vesicles (CCV) mediate protein sorting and vesicular trafficking from the plasma membrane and the trans-Golgi network. Before delivery of the vesicle contents to the target organelles, the coat components, clathrin and adaptor protein complexes (APs), must be released. Previous work has established that hsc70/the uncoating ATPase mediates clathrin release in vitro without the release of APs. AP release has not been reconstituted in vitro, and nothing is known about the requirements for this reaction. We report a novel quantitative assay for the ATP- and cytosol- dependent release of APs from CCV. As expected, hsc70 is not sufficient for AP release; however, immunodepletion and reconstitution experiments establish that it is necessary. Interestingly, complete clathrin release is not a prerequisite for AP release, suggesting that hsc70 plays a dual role in recycling the constituents of the clathrin coat. This assay provides a functional basis for identification of the additional cytosolic factor(s) required for AP release.

MeSH Terms
Adenosine Triphosphate/metabolism Animals Brain/metabolism Carrier Proteins/metabolism Cattle Clathrin/metabolism Coated Pits, Cell-Membrane/metabolism,ultrastructure Cytosol/metabolism Enzyme-Linked Immunosorbent Assay HSC70 Heat-Shock Proteins HSP70 Heat-Shock Proteins Kinetics Membrane Proteins/metabolism
Chemicals
Carrier Proteins Clathrin HSC70 Heat-Shock Proteins HSP70 Heat-Shock Proteins Membrane Proteins Adenosine Triphosphate
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hannan L A
Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
Newmyer S L
Schmid S L
References (54)
54 references, click to expand
  1. In vitro binding of plasma membrane-coated vesicle adaptors to the cytoplasmic domain of lysosomal acid phosphatase.
    J Biol Chem. 1993 Jun 15;268(17):12537-43 PMID: 8509395
  2. Association of the AP-3 adaptor complex with clathrin.
    Science. 1998 Apr 17;280(5362):431-4 PMID: 9545220
  3. Nucleotide binding properties of bovine brain uncoating ATPase.
    J Biol Chem. 1993 Apr 25;268(12):8507-13 PMID: 8473294
  4. Coated vesicles: characterization, selective dissociation, and reassembly.
    Proc Natl Acad Sci U S A. 1978 Sep;75(9):4394-8 PMID: 30086
  5. Preparation of membrane-depleted polygonal coat structures from isolated coated vesicles.
    Cell Biol Int Rep. 1978 Sep;2(5):457-64 PMID: 709607
  6. Clathrin-coated vesicles: isolation, dissociation and factor-dependent reassociation of clathrin baskets.
    Cell. 1979 Feb;16(2):303-12 PMID: 455437
  7. Assembly and packing of clathrin into coats.
    J Cell Biol. 1981 Dec;91(3 Pt 1):790-7 PMID: 7328122
  8. Coated vesicles from human placenta carry ferritin, transferrin, and immunoglobulin G.
    Proc Natl Acad Sci U S A. 1982 Jan;79(2):451-5 PMID: 6952195
  9. An enzyme that removes clathrin coats: purification of an uncoating ATPase.
    J Cell Biol. 1984 Aug;99(2):723-33 PMID: 6146630
  10. Dissociation of clathrin coats coupled to the hydrolysis of ATP: role of an uncoating ATPase.
    J Cell Biol. 1984 Aug;99(2):734-41 PMID: 6146631
  11. Purification and properties of 100-kd proteins from coated vesicles and their reconstitution with clathrin.
    EMBO J. 1984 Sep;3(9):1951-7 PMID: 6149117
  12. Assembled and unassembled pools of clathrin: a quantitative study using an enzyme immunoassay.
    J Cell Biol. 1985 Feb;100(2):521-7 PMID: 3968176
  13. Two classes of binding sites for uncoating protein in clathrin triskelions.
    J Biol Chem. 1985 Aug 25;260(18):10050-6 PMID: 2862147
  14. Three-dimensional structure of clathrin cages in ice.
    EMBO J. 1986 Mar;5(3):529-34 PMID: 3635476
  15. Enzymatic recycling of clathrin from coated vesicles.
    Cell. 1986 Jul 4;46(1):5-9 PMID: 2872968
  16. Location of the 100 kd-50 kd accessory proteins in clathrin coats.
    EMBO J. 1986 Sep;5(9):2079-85 PMID: 2877872
  17. 100-kD coated vesicle proteins: molecular heterogeneity and intracellular distribution studied with monoclonal antibodies.
    J Cell Biol. 1987 Apr;104(4):887-95 PMID: 2881934
  18. Structural relationships between clathrin assembly proteins from the Golgi and the plasma membrane.
    EMBO J. 1988 Apr;7(4):919-29 PMID: 3402440
  19. Deep-etch visualization of proteins involved in clathrin assembly.
    J Cell Biol. 1988 Sep;107(3):877-86 PMID: 3417785
  20. Receptors compete for adaptors found in plasma membrane coated pits.
    EMBO J. 1988 Nov;7(11):3331-6 PMID: 2905261
  21. Trimeric binding of the 70-kD uncoating ATPase to the vertices of clathrin triskelia: a candidate intermediate in the vesicle uncoating reaction.
    J Cell Biol. 1989 Oct;109(4 Pt 1):1457-66 PMID: 2571614
  22. Identification of a clathrin binding subunit in the HA2 adaptor protein complex.
    J Biol Chem. 1989 Nov 25;264(33):20089-93 PMID: 2573598
  23. 100-kDa polypeptides in peripheral clathrin-coated vesicles are required for receptor-mediated endocytosis.
    Proc Natl Acad Sci U S A. 1989 Dec;86(23):9289-93 PMID: 2574457
  24. Dissociation of clathrin from coated vesicles by the uncoating ATPase.
    J Biol Chem. 1990 Apr 25;265(12):6682-7 PMID: 1969864
  25. Auxilin, a newly identified clathrin-associated protein in coated vesicles from bovine brain.
    J Cell Biol. 1990 Jul;111(1):19-29 PMID: 1973169
  26. Clathrin-coated pits contain an integral membrane protein that binds the AP-2 subunit with high affinity.
    J Biol Chem. 1990 Sep 25;265(27):16514-20 PMID: 1975814
  27. Stabilization of clathrin coats by the core of the clathrin-associated protein complex AP-2.
    Biochemistry. 1990 Dec 4;29(48):10791-8 PMID: 2125494
  28. Evidence that the 90-kDa heat shock protein (HSP90) exists in cytosol in heteromeric complexes containing HSP70 and three other proteins with Mr of 63,000, 56,000, and 50,000.
    J Biol Chem. 1991 Apr 15;266(11):6708-13 PMID: 2016286
  29. Subunit interaction and function of clathrin-coated vesicle adaptors from the Golgi and the plasma membrane.
    J Biol Chem. 1991 Apr 25;266(12):7910-8 PMID: 1902231
  30. Clathrin domains involved in recognition by assembly protein AP-2.
    J Biol Chem. 1991 Apr 25;266(12):7950-6 PMID: 1902233
  31. Interaction of assembly protein AP-2 and its isolated subunits with clathrin.
    Biochemistry. 1991 Jun 4;30(22):5590-7 PMID: 2036430
  32. Clathrin assembly protein AP-2 induces aggregation of membrane vesicles: a possible role for AP-2 in endosome formation.
    J Cell Biol. 1992 Nov;119(4):787-96 PMID: 1358896
  33. Complex formation between clathrin and uncoating ATPase.
    J Biol Chem. 1994 Mar 4;269(9):6931-9 PMID: 8120055
  34. Molecular chaperones in protein folding: the art of avoiding sticky situations.
    Trends Biochem Sci. 1994 Jan;19(1):20-5 PMID: 7908149
  35. Characterization of the protein-protein interactions determining the heat shock protein (hsp90.hsp70.hsp56) heterocomplex.
    J Biol Chem. 1994 Apr 15;269(15):11155-61 PMID: 8157642
  36. DnaJ-like proteins: molecular chaperones and specific regulators of Hsp70.
    Trends Biochem Sci. 1994 Apr;19(4):176-81 PMID: 8016869
  37. Receptor-mediated endocytosis is sensitive to antibodies against the uncoating ATPase (hsc70).
    J Cell Sci. 1994 May;107 ( Pt 5):1185-96 PMID: 7929628
  38. The role of clathrin, adaptors and dynamin in endocytosis.
    Curr Opin Cell Biol. 1994 Aug;6(4):538-44 PMID: 7986531
  39. Different domains of the AP-1 adaptor complex are required for Golgi membrane binding and clathrin recruitment.
    J Biol Chem. 1995 Mar 3;270(9):4933-42 PMID: 7876268
  40. Selective action of uncoating ATPase towards clathrin-coated vesicles from brain.
    J Cell Sci. 1995 Mar;108 ( Pt 3):1295-306 PMID: 7622612
  41. Interaction of tyrosine-based sorting signals with clathrin-associated proteins.
    Science. 1995 Sep 29;269(5232):1872-5 PMID: 7569928
  42. Expression and localization of alpha-adaptin isoforms.
    J Cell Sci. 1995 Aug;108 ( Pt 8):2865-75 PMID: 7593326
  43. Coated vesicles: a diversity of form and function.
    FASEB J. 1995 Nov;9(14):1445-53 PMID: 7589986
  44. Role of auxilin in uncoating clathrin-coated vesicles.
    Nature. 1995 Dec 7;378(6557):632-5 PMID: 8524399
  45. Coat proteins and vesicle budding.
    Science. 1996 Mar 15;271(5255):1526-33 PMID: 8599108
  46. Protein sorting by transport vesicles.
    Science. 1996 Apr 12;272(5259):227-34 PMID: 8602507
  47. Purification and characterization of a 66-kDa protein from rabbit reticulocyte lysate which promotes the recycling of hsp 70.
    J Biol Chem. 1996 Jul 12;271(28):16833-41 PMID: 8663320
  48. Binding of ATP and ATP analogues to the uncoating ATPase Hsc70 (70 kDa heat-shock cognate protein).
    Biochem J. 1996 Sep 15;318 ( Pt 3):923-9 PMID: 8836139
  49. In vivo phosphorylation of adaptors regulates their interaction with clathrin.
    J Cell Biol. 1996 Nov;135(3):635-45 PMID: 8909539
  50. Characterization of the adaptor-related protein complex, AP-3.
    J Cell Biol. 1997 May 19;137(4):835-45 PMID: 9151686
  51. Steroid receptor interactions with heat shock protein and immunophilin chaperones.
    Endocr Rev. 1997 Jun;18(3):306-60 PMID: 9183567
  52. Clathrin-coated vesicle formation and protein sorting: an integrated process.
    Annu Rev Biochem. 1997;66:511-48 PMID: 9242916
  53. Linking cargo to vesicle formation: receptor tail interactions with coat proteins.
    Curr Opin Cell Biol. 1997 Aug;9(4):488-95 PMID: 9261055
  54. Multiple GTP-binding proteins participate in clathrin-coated vesicle-mediated endocytosis.
    J Cell Biol. 1993 Jan;120(1):37-45 PMID: 8416994
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.9.8.2217
Published
1998-08-00
Pages
2217-29
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC25474
Subset
IM
Grants
NCI NIH HHS · CA-58689 · United States
NIGMS NIH HHS · GM-42455 · United States
External Links
PubMed source DOI: 10.1091/mbc.9.8.2217 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2