Release of both native and non-native proteins from a cis-only GroEL ternary complex.
Protein folding by the double-ring chaperonin GroEL is initiated in cis ternary complexes, in which polypeptide is sequestered in the central channel of a GroEL ring, capped by the co-chaperonin GroES. The cis ternary complex is dissociated (half-life of approximately 15 s) by trans-sided ATP hydrolysis, which triggers release of GroES. For the substrate protein rhodanese, only approximately 15% of cis-localized molecules attain their native form before hydrolysis. A major question concerning the GroEL mechanism is whether both native and non-native forms are released from the cis complex. Here we address this question using a 'cis-only' mixed-ring GroEL complex that binds polypeptide and GroES on only one of its two rings. This complex mediates refolding of rhodanese but, as with wild-type GroEL, renaturation is quenched by addition of mutant GroEL 'traps', which bind but do not release polypeptide substrate. This indicates that non-native forms are released from the cis complex. Quenching of refolding by traps was also observed under physiological conditions, both in undiluted Xenopus oocyte extract and in intact oocytes. We conclude that release of non-native forms from GroEL in vivo allows a kinetic partitioning among various chaperones and proteolytic components, which determines both the conformation and lifetime of a protein.
Burston
S G
SG
Department of Genetics and Howard Hughes Medical Institute, New Haven, Connecticut 06510, USA.
Weissman
J S
JS
Farr
G W
GW
Fenton
W A
WA
Horwich
A L
AL
D000244:Adenosine Diphosphate / Q000235:genetics / Q000378:metabolism; D000818:Animals; D018835:Chaperonin 10 / Q000235:genetics / Q000378:metabolism; D018834:Chaperonin 60 / Q000235:genetics / Q000378:metabolism*; D046911:Macromolecular Substances; D011485:Protein Binding; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013884:Thiosulfate Sulfurtransferase / Q000378:metabolism; D014981:Xenopus
D018835:Chaperonin 10; D018834:Chaperonin 60; D046911:Macromolecular Substances; D011993:Recombinant Fusion Proteins; D000244:Adenosine Diphosphate; D013884:Thiosulfate Sulfurtransferase