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PMID: 2182655 Published · ppublish English Journal Article

Identification of a 25-kD protein from yeast cytosol that operates in a prefusion step of vesicular transport between compartments of the Golgi.

The Journal of cell biology ·Vol. 110 ·No. 4 ·1990-04-00 ·Pages 947-54

Wattenberg BW, Hiebsch RR, LeCureux LW, White MP

Abstract

We have identified a 25-kD cytosolic yeast protein that mediates a late, prefusion step in transport of proteins between compartments of the Golgi apparatus. Activity was followed using the previously described cell free assay for protein transport between Golgi compartments as modified to detect late acting cytosolic factors (Wattenberg, B. W., and J. E. Rothman. 1986. J. Biol. Chem. 263:2208-2213). In the reaction mediated by this protein, transport vesicles that have become attached to the target membrane during a preincubation are processed in preparation for fusion. The ultimate fusion event does not require the addition of cytosolic proteins (Balch, W. E., W. G. Dunphy, W. A. Braell, and J. E. Rothman. 1984. Cell. 39:525-536). Although isolated from yeast, this protein has activity when assayed with mammalian membranes. This protein has been enriched over 150-fold from yeast cytosol, albeit not to complete homogeneity. The identity of a 25-kD polypeptide as the active component was confirmed by raising monoclonal antibodies to it. These antibodies were found to specifically inhibit transport activity. Because this is a protein operating in prefusion, it has been abbreviated POP.

MeSH Terms
Animals Antibodies, Monoclonal Biological Transport Cell Line Chromatography, Gel Chromatography, Ion Exchange Cytosol/metabolism Electrophoresis, Polyacrylamide Gel Fungal Proteins/isolation & purification,metabolism Golgi Apparatus/metabolism Immunoblotting Kinetics Molecular Weight Organelles/metabolism Saccharomyces cerevisiae/metabolism
Chemicals
Antibodies, Monoclonal Fungal Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Wattenberg B W
Cell Biology Unit, Upjohn Company, Kalamazoo, Michigan 49001.
Hiebsch R R
LeCureux L W
White M P
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.110.4.947
Published
1990-04-00
Pages
947-54
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2116070
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.110.4.947 PMC Full Text View journal details
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