Reactivation and refolding of a partially folded creatine kinase modified by 5,5'-dithio-bis(2-nitrobenzoic acid).
Creatine kinase with its thiol groups modified by 5, 5'-dithio-bis(2-nitrobenzoic acid) has been shown to be partially folded in a monomeric state using fluorescence, circular dichroism, proteolysis, and size exclusion chromatography studies. In the presence of DTT, the partially folded modified creatine kinase can be reactivated and refolded following a biphasic course, suggesting the existence of a monomeric intermediate during the refolding of CK. The results provide evidence for our previously suggested model of the refolding pathway of urea-denatured creatine kinase.
Yang
Y
Y
Department of Biological Science and Biotechnology, School of Life Science and Engineering, Beijing, 100084, China.
Park
Y D
YD
Yu
T W
TW
Zhou
H M
HM
D000818:Animals; D002942:Circular Dichroism; D003402:Creatine Kinase / Q000737:chemistry*; D004228:Dithionitrobenzoic Acid / Q000494:pharmacology*; D004229:Dithiothreitol / Q000737:chemistry; D004789:Enzyme Activation; D007700:Kinetics; D018482:Muscle, Skeletal / Q000201:enzymology; D017510:Protein Folding; D011817:Rabbits; D013050:Spectrometry, Fluorescence; D013438:Sulfhydryl Compounds / Q000737:chemistry
D013438:Sulfhydryl Compounds; D004228:Dithionitrobenzoic Acid; D003402:Creatine Kinase; D004229:Dithiothreitol