PMID: 11311859Apr 20, 2001Paper

Aggregation of creatine kinase during refolding and chaperonin-mediated folding of creatine kinase

The International Journal of Biochemistry & Cell Biology
S LiH Zhou

Abstract

The course of refolding and reactivation of urea-denatured creatine kinase (ATP; creatine N-phosphotransferase, EC 2.7.3.2) has been studied in the absence and presence of molecular chaperonin GroEL. The enzyme was denatured in Tris--HCl buffer containing 6 M urea for 1 h. In the refolding studies, the denatured enzyme was diluted 60-fold into the same buffer containing GroEL or not for activity, turbidity, fluorescence measurements and polyacrylamide gel electrophoresis. The results show that the reactivation process is dependent of creatine kinase concentration in the concentration range 2.5--4 microM. The levels of activity recovery decrease with increasing enzyme concentration because of the formation of wrong aggregates. The molecular chaperonin GroEL can bind the refolding intermediate of creatine kinase and thus prevent the formation of wrong aggregates. This intermediate is an inactive dimeric form that is in a conformation resembling the 'molten globule' state.

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Citations

Jun 13, 2008·Applied Biochemistry and Biotechnology·He-Chang ZouYong-Doo Park
Jan 26, 2002·The International Journal of Biochemistry & Cell Biology·Wen-Bin OuHai-Meng Zhou
May 15, 2002·The International Journal of Biochemistry & Cell Biology·Wen bin OuHai Meng Zhou
Jun 27, 2013·Protein Engineering, Design & Selection : PEDS·Sen LiYixin Zhang
Oct 18, 2001·Protein Science : a Publication of the Protein Society·W B OuH M Zhou
Aug 19, 2010·International Journal of Molecular Sciences·Yan-Song GaoYong-Bin Yan
May 11, 2013·Protein Science : a Publication of the Protein Society·Shan FengHaiteng Deng
Oct 13, 2006·The International Journal of Biochemistry & Cell Biology·Shan FengYong-Bin Yan
May 2, 2012·International Journal of Biological Macromolecules·Yong-Qiang FanYu-Long Wang
Aug 14, 2012·International Journal of Biological Macromolecules·Yong-Qiang FanYu-Long Wang
Aug 27, 2016·PloS One·Nevena HristozovaDenes Kovacs
Mar 10, 2010·The Protein Journal·Akiko FujimotoKentaro Shiraki
Aug 31, 2001·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Y D ParkH M Zhou
Sep 18, 2002·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Wen-Bin OuHai-Meng Zhou

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