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PMID: 9482852 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Reversible dissociation of coatomer: functional characterization of a beta/delta-coat protein subcomplex.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 95 ·No. 5 ·1998-03-03 ·Pages 2140-5

Pavel J, Harter C, Wieland FT

Abstract

COPI-coated vesicles mediate protein transport within the early secretory pathway. Their coat consists of ADP ribosylation factor (ARF1, a small guanosine nucleotide binding protein), and coatomer, a cytosolic complex composed of seven subunits, alpha- to zeta-coat proteins (COPs). For coat formation that initiates budding of a vesicle, ARF1 is recruited to the Golgi membrane from the cytosol in its GTP-bound form, and subsequently, coatomer can bind to the membrane. To identify a minimal structure of coatomer capable to bind to Golgi membranes in an ARF1-dependent manner, we have established a procedure to dissociate coatomer under conditions that allow reassociation of the subunits to a complete and functional complex. After dissociation, subunits or subcomplexes can be isolated and may be expected to be functional. Herein we describe isolation of a subcomplex of coatomer consisting of beta- and delta-COPs that is able to bind to Golgi membranes in an ARF1- and GTP-dependent manner.

MeSH Terms
ADP-Ribosylation Factor 1 ADP-Ribosylation Factors Animals Coatomer Protein Cytosol/metabolism Dimerization GTP-Binding Proteins/metabolism Golgi Apparatus/metabolism Guanosine 5'-O-(3-Thiotriphosphate)/pharmacology Guanosine Diphosphate/analogs & derivatives,pharmacology Guanosine Triphosphate/metabolism Liver/metabolism Macromolecular Substances Maleic Anhydrides/pharmacology Membrane Proteins/chemistry,isolation & purification,metabolism Microtubule-Associated Proteins/isolation & purification,metabolism Rabbits Thionucleotides/pharmacology
Chemicals
Coatomer Protein Macromolecular Substances Maleic Anhydrides Membrane Proteins Microtubule-Associated Proteins Thionucleotides Guanosine Diphosphate Guanosine 5'-O-(3-Thiotriphosphate) 2,3-dimethylmaleic anhydride guanosine 5'-O-(2-thiodiphosphate) Guanosine Triphosphate GTP-Binding Proteins ADP-Ribosylation Factor 1 ADP-Ribosylation Factors
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Pavel J
Biochemie-Zentrum Heidelberg Ruprecht-Karls-Universität, Im Neuenheimer Feld 328, 69120 Heidelberg, Germany.
Harter C
Wieland F T
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.95.5.2140
Published
1998-03-03
Pages
2140-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC19276
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.95.5.2140 PMC Full Text View journal details
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