Protein disulfide isomerase-catalyzed renaturation of ribonuclease A modified by S-thiolation with glutathione and cysteine.
Renaturation of modified ribonuclease A by protein disulfide isomerase was studied. The renaturation rate of fully S-thiolated ribonuclease A with glutathione, namely, ribonuclease A-glutathione mixed disulfide (RNase-SG) containing 8 moles of glutathione per mole of ribonuclease A (RNase-SG8), by protein disulfied isomerase (PDI) was more than three times faster than those of fully S-thiolated RNase with L-cysteine and scrambled ribonuclease A. Renaturation of RNase-SG species containing 7 or less glutathione was slower than that of RNase-SG8. These data seems to favor the hypothesis that S-thiolation of nascent proteins with glutathione may occur in the folding process during protein synthesis. The applicability of the present method consisted of chemical S-thiolation and PDI-catalyzed renaturation to the in vitro folding of recombinant cysteine-containing proteins is discussed.
Ubuka
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Department of Biochemistry, Okayama University Medical School, Japan.
D000818:Animals; D002417:Cattle; D003545:Cysteine / Q000378:metabolism* / Q000494:pharmacology*; D005978:Glutathione / Q000378:metabolism* / Q000494:pharmacology*; D007535:Isomerases / Q000378:metabolism*; D007700:Kinetics; D011489:Protein Denaturation; D019704:Protein Disulfide-Isomerases; D017510:Protein Folding; D012259:Ribonuclease, Pancreatic / Q000737:chemistry / Q000378:metabolism*; D013053:Spectrophotometry; D013438:Sulfhydryl Compounds / Q000378:metabolism
D013438:Sulfhydryl Compounds; D012259:Ribonuclease, Pancreatic; D007535:Isomerases; D019704:Protein Disulfide-Isomerases; D005978:Glutathione; D003545:Cysteine