title: Biochemical heterogeneity and phosphorylation of coatomer subunits.
abstract: The coat protomer complex I (COPI) family of coat proteins are involved in the assembly of membrane-associated coats thought to mediate vesicular transport between the endoplasmic reticulum and the Golgi complex, between adjacent Golgi cisternae, and possibly in the endocytic pathway. We investigated whether this heterogeneity in the sites of COPI action might be reflected in biochemical heterogeneity of one or more COPI subunits. A simplified method was devised to purify the cytosolic COPI precursor complex, coatomer, from rat liver cytosol. The individual subunits were analyzed by high resolution two dimensional gel electrophoresis and mass spectroscopic analysis of tryptic peptides. Considerable charge heterogeneity was observed, particularly for the beta-COP and delta-COP subunits. The multiple species detected, however, did not appear to reflect the presence of distinct translation products but rather a significant degree of protein phosphorylation. The observed pI of beta-COP was sensitive to alkaline phosphatase digestion. Moreover, isolation of coatomer from metabolically labeled tissue culture cells demonstrated directly that both beta-COP and delta-COP, but no other coatomer subunits, were serine-phosphorylated. COPI phosphorylation may regulate coatomer assembly, membrane recruitment, or the specificity of coatomer-organelle interaction.
authors.affiliation: Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520-8002, USA.
authors.forename: D
authors.initials: D
authors.lastname: Sheff
authors.forename: M
authors.initials: M
authors.lastname: Lowe
authors.forename: T E
authors.initials: TE
authors.lastname: Kreis
authors.forename: I
authors.initials: I
authors.lastname: Mellman
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D006367:HeLa Cells; D006801:Humans; D008099:Liver / Q000378:metabolism; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism*; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D008565:Membrane Proteins