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PMID: 8947548 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A major transmembrane protein of Golgi-derived COPI-coated vesicles involved in coatomer binding.

The Journal of cell biology ·Vol. 135 ·No. 5 ·1996-12-00 ·Pages 1239-48

Sohn K, Orci L, Ravazzola M, Amherdt M, Bremser M, Lottspeich F, Fiedler K, Helms JB, Wieland FT

Abstract

Formation of non-clathrin-coated vesicles requires the recruitment of several cytosolic factors to the Golgi membrane. To identify membrane proteins involved in this budding process, a highly abundant type I transmembrane protein (p23) was isolated from mammalian Golgi-derived COPI-coated vesicles, and its cDNA was cloned and sequenced. It belongs to the p24 family of proteins involved in the budding of transport vesicles (Stamnes, M.A., M.W. Craighead, M.H. Hoe, N. Lampen, S. Geromanos, P. Tempst, and J.E. Rothman. 1995. Proc. Natl. Acad. Sci. USA. 92:8011-8015). p23 consists of a large NH2-terminal luminal domain and a short COOH-terminal cytoplasmic tail (-LRRFFKAKKLIE-CO2-) that shows similarity, but not identity, with the sequence motif-KKXX-CO2-, known as a signal for retrieval of escaped ER-resident membrane proteins (Jackson, M.R., T. Nilsson, and P.A. Peterson. 1990. EMBO (Eur. Mol. Biol. Organ.) J. 9:3153-3162; Nilsson, T., M. Jackson, and P.A. Peterson. 1989. Cell. 58:707-718). The cytoplasmic tail of p23 binds to coatomer with similar efficiency as known KKXX motifs. However, the p23 tail differs from the KKXX motif in having an additional motif needed for binding of coatomer. p23 is localized to Golgi cisternae and, during vesicle formation, it concentrates into COPI-coated buds and vesicles. Biochemical analysis revealed that p23 is enriched in vesicles by a factor of approximately 20, as compared with the donor Golgi fraction, and is present in amounts stoichiometric to the small GTP-binding protein ADP-ribosylation factor (ARF) and coatomer. From these data we conclude that p23 represents a Golgi-specific receptor for coatomer involved in the formation of COPI-coated vesicles.

MeSH Terms
Amino Acid Sequence Animals CHO Cells Cloning, Molecular Coated Vesicles/chemistry,metabolism Coatomer Protein Cricetinae DNA, Complementary/genetics Fluorescent Antibody Technique Golgi Apparatus/chemistry,metabolism Membrane Proteins/analysis,chemistry,isolation & purification,metabolism Microscopy, Immunoelectron Molecular Sequence Data Molecular Weight Receptors, Cytoplasmic and Nuclear
Chemicals
BHKp23 protein, mammalian Coatomer Protein DNA, Complementary Membrane Proteins Receptors, Cytoplasmic and Nuclear
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Sohn K
Institut für Biochemie I, Universität Heidelberg, Germany.
Orci L
Ravazzola M
Amherdt M
Bremser M
Lottspeich F
Fiedler K
Helms J B
Wieland F T
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.135.5.1239
Published
1996-12-00
Pages
1239-48
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2121093
Subset
IM
Databases
GENBANK
X98303
External Links
PubMed source DOI: 10.1083/jcb.135.5.1239 PMC Full Text View journal details
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