GroEL and GroES increase the specific enzymatic activity of newly-synthesized rhodanese if present during in vitro transcription/translation.
Enzymatically active mammalian rhodanese, a mitochondrial matrix enzyme, which has been found to require assistants for efficient refolding in vitro, has been synthesized from a plasmid in a cell-free, fractionated, coupled transcription/translation system derived from Escherichia coli. The bacterial chaperonins, GroEL and GroES, along with the rhodanese substrate thiosulfate greatly enhance the specific enzymatic activity of the rhodanese polypeptide that is formed. Indirect evidence suggests that the effect of the GroEL/ES chaperonins is on ribosome-bound nascent peptides. The in vitro transcription/translation system produces sufficient amounts of rhodanese to provide a system for studying factors that control the initial steps in folding of nascent proteins.
Tsalkova
T
T
Department of Chemistry and Biochemistry, University of Texas, Austin 78712.
Zardeneta
G
G
Kudlicki
W
W
Kramer
G
G
Horowitz
P M
PM
Hardesty
B
B
D001426:Bacterial Proteins / Q000378:metabolism*; D018835:Chaperonin 10; D018834:Chaperonin 60; D018833:Chaperonins; D006360:Heat-Shock Proteins / Q000378:metabolism*; D066298:In Vitro Techniques; D014176:Protein Biosynthesis; D011487:Protein Conformation; D011506:Proteins / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013884:Thiosulfate Sulfurtransferase / Q000378:metabolism* / Q000648:ultrastructure; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D018835:Chaperonin 10; D018834:Chaperonin 60; D006360:Heat-Shock Proteins; D011506:Proteins; D011994:Recombinant Proteins; D013884:Thiosulfate Sulfurtransferase; D018833:Chaperonins