Purification and properties of rat liver Sec23-Sec24 complex.
We have demonstrated a protocol for purifying functional Sec23-24 complex from rat liver cytosol. Because the rat liver Sec23-24 complex is highly susceptible to proteolysis, we have noted several modifications which have allowed us to overcome the problem of degradation. If care is taken to prevent proteolysis, this procedure typically yields 2 mg of functional complex. The Sec23-24 complex can then be used to study Sar1 GTP hydrolysis in the Sec23 GTPase activation assay. Additionally, Sec23-24 can reconstitute ER vesicle formation in the presence of Sar1 and Sec13-31, allowing for the identification of novel proteins or compounds that affect cargo export.
Weissman
J T
JT
Department of Cell Biology, Scripps Research Institute, La Jolla, California 92037, USA.
Aridor
M
M
Balch
W E
WE
D000818:Animals; D022181:COP-Coated Vesicles / Q000378:metabolism; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004789:Enzyme Activation; D007700:Kinetics; D008099:Liver / Q000737:chemistry*; D046911:Macromolecular Substances; D008297:Male; D008565:Membrane Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D020559:Monomeric GTP-Binding Proteins / Q000378:metabolism; D011506:Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D029701:Saccharomyces cerevisiae Proteins; D033921:Vesicular Transport Proteins
D046911:Macromolecular Substances; D008565:Membrane Proteins; D011506:Proteins; C118289:SEC24 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C512740:Sec23a protein, rat; D033921:Vesicular Transport Proteins; D020559:Monomeric GTP-Binding Proteins; C077395:SAR1 protein, S cerevisiae