caa a22 4500 39755558X CHVBK 20180308164801.0 cr unu---uuuuu 161202e199612 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021528 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021528 Effects of Subtilisin Cleavage of Monomeric Actin on Its Nucleotide Binding [Elektronische Daten] [Atsushi Ooi, Koshin Mihashi] The kinetics of ATP exchange on subtilisin-cleaved G-actin was investigated by measuring the fluorescence of 1,N6-ethenoadenosine 5'-triphosphate. The apparent dissociation rate of ATP (k−ATP) was 2.8-fold larger than that of intact G-actin in the presence of 300 μM free Ca2+. Analysis of the dependence of Kk−ATP on free Ca2+ showed that the dissociation rate constant of tightly bound Ca2+ was not significantly changed by subtilisin cleavage. On the other hand, an equilibrium binding study using 8-amino-2-[(2-amino-5-methylphenoxy)-methyl]-6-methoxyquinoline N,N,N',N'-tetraacetic acid (Quin 2) showed that the affinity of tightly bound Ca2+ for G-actin was reduced by about 13-fold after subtilisin treatment. Consequently, the stabilization by Ca2+ of ATP was weak in cleaved G-actin. Furthermore, the kinetic analysis of ATP exchange revealed that the binding equilibrium between ATP and divalent cation-free cleaved G-actin was much slower than that in the case of intact G-actin. © 1996 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence actin nationallicence divalent cation nationallicence nucleotide nationallicence subtilisin nationallicence surface loop nationallicence Ooi Atsushi Faculty of Bioresources, Mie University, Kamihama 1515, Tsu, 514, Furo-cho, Chikusa-ku, Nagoya 465-01 aut Mihashi Koshin Faculty of Bioresources, Mie University Kamihama 1515, Tsu, 514 aut The Journal of Biochemistry Oxford University Press 120/6(1996-12), 1104-1110 0021-924X 120:6<1104 1996 120 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021528 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021528 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ooi Atsushi Faculty of Bioresources, Mie University, Kamihama 1515, Tsu, 514, Furo-cho, Chikusa-ku, Nagoya 465-01 aut NATIONALLICENCE 700 1- Mihashi Koshin Faculty of Bioresources, Mie University Kamihama 1515, Tsu, 514 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 120/6(1996-12), 1104-1110 0021-924X 120:6<1104 1996 120 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford