Purification and properties of mammalian Sec23/24 from insect cells.
The Sec23/24 complex is a large heterodimeric protein involved in COPII vesicle biogenesis. The individual mammalian protein subunits are too large for expression in bacterial systems. This article details the use of the Bac-to-Bac baculovirus coexpression system in insect cells for both the human Sec23A and Sec24C. This strategy results in high yields of pure, functional protein and can be adapted for the purification of other Sec23/24 isoforms for their biochemical and biological characterization.
LaPointe
Paul
P
The Scripps Research Institute, Department of Cell Biology, La Jolla, California, USA.
Balch
William E
WE
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D022181:COP-Coated Vesicles / Q000201:enzymology; D002846:Chromatography, Affinity / Q000379:methods; D006801:Humans; D020559:Monomeric GTP-Binding Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification; D018411:Spodoptera; D033921:Vesicular Transport Proteins / Q000302:isolation & purification*
D011994:Recombinant Proteins; C512739:SEC23A protein, human; C530640:SEC24C protein, human; D033921:Vesicular Transport Proteins; C493083:SAR1A protein, human; C513282:SAR1B protein, human; D020559:Monomeric GTP-Binding Proteins