Mammalian cytosolic chaperonin.
Cytosolic chaperonin, the eukaryotic cytosolic homolog of GroEL, has certain unusual features that make it uniquely useful for studying the mechanism of chaperonin action. It is of particular interest as an essential component in the generation of native actin and tubulin in vivo. We describe a method for the purification of mammalian c-cpn from rabbit reticulocyte lysate via a three-step procedure involving ion-exchange chromatography, affinity selection on ATP-agarose, and gel filtration. We also describe a sensitive in vitro-folding assay for the activity of c-cpn and other chaperone proteins, and a simple nondenaturing gel assay for the analysis of folding reaction products.
Cowan
N J
NJ
Department of Biochemistry, New York University School of Medicine, New York 10016-6402, USA.
D000199:Actins / Q000378:metabolism; D000818:Animals; D018834:Chaperonin 60 / Q000737:chemistry; D056404:Chaperonin Containing TCP-1; D018833:Chaperonins / Q000737:chemistry* / Q000378:metabolism; D003600:Cytosol / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D011487:Protein Conformation; D017510:Protein Folding; D011817:Rabbits; D012156:Reticulocytes / Q000737:chemistry; D014404:Tubulin / Q000378:metabolism
D000199:Actins; D018834:Chaperonin 60; D014404:Tubulin; D056404:Chaperonin Containing TCP-1; D018833:Chaperonins