Purification of an N-ethylmaleimide-Sensitive Protein Catalyzing Vesicular Transport

N-Ethylmaleimide (NEM) inhibits protein transport between successive compartments of the Golgi stack in a cell-free system. After inactivation of the Golgi membranes by NEM, transport can be rescued by adding back an appropriately prepared cytosol fraction. This complementation assay has allowed us...

Full description

Saved in:
Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1988-11, Vol.85 (21), p.7852-7856
Main Authors: Block, Marc R., Glick, Benjamin S., Wilcox, Celeste A., Wieland, Felix T., Rothman, James E.
Format: Article
Language:English
Subjects:
Adenosine Triphosphate - metabolism
Analytical, structural and metabolic biochemistry
Animals
Antibodies
Antibodies, Monoclonal
Binding and carrier proteins
Biochemistry
Biological and medical sciences
Biological Transport, Active - drug effects
Carrier Proteins - isolation & purification
Carrier Proteins - pharmacology
Cell Line
Cell physiology
CHO cells
Cricetinae
Cytosol
Cytosol - analysis
Fundamental and applied biological sciences. Psychology
Gels
Golgi apparatus
Golgi Apparatus - metabolism
Homogenization
Ice
Membrane and intracellular transports
Molecular and cellular biology
Monoclonal antibodies
Proteins
Transport vesicles
vesicles
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:N-Ethylmaleimide (NEM) inhibits protein transport between successive compartments of the Golgi stack in a cell-free system. After inactivation of the Golgi membranes by NEM, transport can be rescued by adding back an appropriately prepared cytosol fraction. This complementation assay has allowed us to purify the NEM-sensitive factor, which we term NSF. The NEM-sensitive factor is a tetramer of 76-kDa subunits, and appears to act catalytically, one tetramer leading to the metabolism of numerous transport vesicles.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.85.21.7852