Architecture of coatomer: Molecular characterization of δ-COP and protein interactions within the complex Journal Article
| Authors: | Faulstich, D.; Auerbach, S.; Orci, L.; Ravazzola, M.; Wegehingel, S.; Lottspeich, F.; Stenbeck, G.; Harter, C.; Wieland, F. T.; Tschochner, H. |
| Article Title: | Architecture of coatomer: Molecular characterization of δ-COP and protein interactions within the complex |
| Abstract: | Coatomer is a cytosolic protein complex that forms the coat of COP 1- coated transport vesicles. In our attempt to analyze the physical and functional interactions between its seven subunits (coat proteins, [COPs] α- ζ), we engaged in a program to clone and characterize the individual coatomer subunits. We have now cloned, sequenced, and overexpressed bovine α-COP, the 135-kD subunit of coatomer as well as δ-COP, the 57-kD subunit and have identified a yeast homolog of δ-COP by cDNA sequence comparison and by NH2-terminal peptide sequencing. δ-COP shows homologies to subunits of the clathrin adaptor complexes AP1 and AP2. We show that in Golgi-enriched membrane fractions, the protein is predominantly found in COP 1-coated transport vesicles and in the budding regions of the Golgi membranes. A knock-out of the δ-COP gene in yeast is lethal. Immunoprecipitation, as well as analysis exploiting the two hybrid system in a complete COP screen, showed physical interactions between α- and εCOPs and between β-COPs. Moreover, the two-hybrid system indicates interactions between γ- and ζ-COPs as well as between α- and β'-COPs. We propose that these interactions reflect in vivo associations of those subunits and thus play a functional role in the assembly of coatomer and/or serve to maintain the molecular architecture of the complex. |
| Keywords: | nonhuman; animals; gene expression; protein protein interaction; cell protein; membrane proteins; intracellular transport; cloning, molecular; liver; amino acid sequence; molecular sequence data; sequence homology, amino acid; recombinant fusion proteins; brain; protein transport; cattle; rats; dna, complementary; genes, lethal; sequence analysis, dna; golgi complex; bovinae; coatomer protein; coated vesicles; golgi apparatus; bos taurus; precipitin tests; priority journal; article |
| Journal Title: | Journal of Cell Biology |
| Volume: | 135 |
| Issue: | 1 |
| ISSN: | 0021-9525 |
| Publisher: | Rockefeller University Press |
| Date Published: | 1996-10-01 |
| Start Page: | 53 |
| End Page: | 61 |
| Language: | English |
| DOI: | 10.1083/jcb.135.1.53 |
| PUBMED: | 8858162 |
| PROVIDER: | scopus |
| PMCID: | PMC2121028 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 22 November 2017 -- Source: Scopus |
Altmetric
Citation Impact
BMJ Impact Analytics
Related MSK Work