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PMID: 1512287 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A novel 115-kD peripheral membrane protein is required for intercisternal transport in the Golgi stack.

The Journal of cell biology ·Vol. 118 ·No. 5 ·1992-09-00 ·Pages 1015-26

Waters MG, Clary DO, Rothman JE

Abstract

We have used an in vitro Golgi protein transport assay dependent on high molecular weight (greater than 100 kD) cytosolic and/or peripheral membrane proteins to study the requirements for transport from the cis- to the medial-compartment. Fractionation of this system indicates that, besides the NEM-sensitive fusion protein (NSF) and the soluble NSF attachment protein (SNAP), at least three high molecular weight protein fractions from bovine liver cytosol are required. The activity from one of these fractions was purified using an assay that included the second and third fractions in a crude state. The result is a protein of 115-kD subunit molecular mass, which we term p115. Immunodepletion of the 115-kD protein from a purified preparation with mAbs removes activity. Peptide sequence analysis of tryptic peptides indicates that p115 is a "novel" protein that has not been described previously. Gel filtration and sedimentation analysis indicate that, in its native state, p115 is a nonglobular homo-oligomer. p115 is present on purified Golgi membranes and can be extracted with high salt concentration or alkaline pH, indicating that it is peripherally associated with the membrane. Indirect immunofluorescence indicates that p115 is associated with the Golgi apparatus in situ.

MeSH Terms
Animals Antibodies, Monoclonal Biological Transport Carrier Proteins/chemistry,immunology,isolation & purification,metabolism Cattle Cell Line Cytosol/chemistry Golgi Apparatus/chemistry,metabolism Liver/chemistry Membrane Glycoproteins Membrane Proteins/chemistry,immunology,isolation & purification,metabolism Molecular Weight Viral Envelope Proteins/metabolism
Chemicals
Antibodies, Monoclonal Carrier Proteins G protein, vesicular stomatitis virus Membrane Glycoproteins Membrane Proteins Viral Envelope Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Waters M G
Program of Cellular Biochemistry and Biophysics, Sloan-Kettering Institute, New York 10021.
Clary D O
Rothman J E
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.118.5.1015
Published
1992-09-00
Pages
1015-26
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2289595
Subset
IM
Grants
NIDDK NIH HHS · DK27044 · United States
External Links
PubMed source DOI: 10.1083/jcb.118.5.1015 PMC Full Text View journal details
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