caa a22 4500 475744233 CHVBK 20180406123508.0 cr unu---uuuuu 170329e20000401xx s 000 0 eng 10.1023/A:1007003703087 doi (NATIONALLICENCE)springer-10.1023/A:1007003703087 Effect of Mg2+ During Reactivation and Refolding of Guanidine Hydrochloride-Denatured Creatine Kinase [Elektronische Daten] [Yong-Doo Park, Hai-Meng Zhou] Creatine kinase (ATP: creatine N-phosphotransferase, EC 2.7.3.2) was completely denatured using 3 M guanidine hydrochloride for 2 h as in previous studies [Yao et al. (1982), Sci. Sin. 25B, 1296-1302; Yao et al. (1984), Biochemistry 23, 2740-2744; Yao et al. (1982), Sci. Sin. 25B, 1186-1193]. Under suitable conditions, about 60-70% of the activity can be recovered in the presence of different Mg2+ concentrations. Both the reactivation and the refolding processes follow two-phase courses after dilution in the proper solutions. A comparison of the rate constants for the refolding of unfolded creatine kinase with those for the recovery of its catalytic activity at various Mg2+ concentrations shows that these are not synchronized. The reactivity of guanidine hydrochloride-denatured creatine kinase can be inhibited by Mg2+; however, the rates of reactivation are independent of the Mg2+ concentration. In addition, Mg2+ affects the fluorescence intensity, but the rate constants of refolding are independent of Mg2+ concentration. Although the reactivation of GdHCl-denatured creatine kinase is complete about 3 h after dilution with reactivation solutions, the conformational changes during refolding occur in a much slower reaction. Mg2+ can induce complex changes in the relative fluorescence intensity during refolding over a broad range of concentrations. Plenum Publishing Corporation, 2000 Creatine kinase nationallicence magnesium ion nationallicence reactivation nationallicence refolding nationallicence Park Yong-Doo Department of Biological Science and Biotechnology, Tsinghua University, 100084, Beijing, China aut Zhou Hai-Meng Department of Biological Science and Biotechnology, Tsinghua University, 100084, Beijing, China aut Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/3(2000-04-01), 193-198 0277-8033 19:3<193 2000 19 10930 https://doi.org/10.1023/A:1007003703087 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1007003703087 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Park Yong-Doo Department of Biological Science and Biotechnology, Tsinghua University, 100084, Beijing, China aut NATIONALLICENCE 700 1- Zhou Hai-Meng Department of Biological Science and Biotechnology, Tsinghua University, 100084, Beijing, China aut NATIONALLICENCE 773 0- Journal of Protein Chemistry Kluwer Academic Publishers-Plenum Publishers 19/3(2000-04-01), 193-198 0277-8033 19:3<193 2000 19 10930 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer