caa a22 4500 39755592X CHVBK 20180308164802.0 cr unu---uuuuu 161202e199601 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021200 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021200 Maleimidobenzoyl Actin: Its Biochemical Properties and In Vitro Motility [Elektronische Daten] [Tetsu Hozumi, Masao Miki, Sugie Higashi-Fujime] Muscle G-actin treated with the hetero-bifunctional cross-linking reagent m-maleimido-benzoyl-N-hydroxysuccinimide ester (MBS), denoted as MBS-G-actin, is not induced to polymerize into F-actin by salt and myosin subfragment 1 [Bettache, N., Bertrand, R., and Kassab, R. (1989) Proc.Natl.Acad.Sci.USA 86,6028-6032]. However, the addition of salt and phalloidin together allowed MBS-G-actin to polymerize and the resulting polymer (P-MBS-G-actin) could activate the Mg2+-ATPase of S-l [Miki, M.and Hozumi.T.(1991) Biochemistry 30, 5625-5630]. When F-actin was treated with MBS (MBS-F-actin), unlike MBS-G-actin, intercross-links between monomers in F-actin occurred. The MBS-F-actin could activate the Mg2+-ATPase of heavy meromyosin (HMM): its maximum turnover rate, Vmax, was almost the same as that of native F-actin, but the affinity of HMM for MBS-F-actin in the presence of ATP, Km, was about 3 times higher. Electron microscopy showed that both P-MBS-G-actin and MBS-F-actin had the double stranded structures of F-actin and formed the arrowhead structures when combined with HMM. By in vitro motility assay, the sliding velocities of P-MBS-G-actin and MBS-F-actin were found to be slightly slower than that of native F-actin. But the critical concentration of KCl, over which the sliding movement was not observed, for MBS-modified actins was considerably higher than for native F-actin. When MBS-modified actins were regulated by tropomyosin-troponin complex, they were less sensitive to the Ca2+ concentration for HMM ATPase activation and sliding movement. These results showed that the modification of some of the lysine residues in the actin molecule leads to change in the biochemical properties of F-actin. © by the Journal of Biochemistry Regular Papers nationallicence actin-actin interface nationallicence actin-myosin interaction nationallicence Ca2+-regulation nationallicence hetero-bifunctional reagent nationallicence in vitro motility assay nationallicence Hozumi Tetsu Department of Physiology, Nagoya City University Medical School Mizuho-ku, Nagoya, Aichi 467 aut Miki Masao Muscle Research Unit, Department of Anatomy, The University of Sydney New South Wales 2006, Australia aut Higashi-Fujime Sugie Department of Molecular Biology, Faculty of Science, Nagoya University Chikusa-ku, Nagoya, Aichi 464 aut The Journal of Biochemistry Oxford University Press 119/1(1996-01), 151-156 0021-924X 119:1<151 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021200 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021200 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hozumi Tetsu Department of Physiology, Nagoya City University Medical School Mizuho-ku, Nagoya, Aichi 467 aut NATIONALLICENCE 700 1- Miki Masao Muscle Research Unit, Department of Anatomy, The University of Sydney New South Wales 2006, Australia aut NATIONALLICENCE 700 1- Higashi-Fujime Sugie Department of Molecular Biology, Faculty of Science, Nagoya University Chikusa-ku, Nagoya, Aichi 464 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/1(1996-01), 151-156 0021-924X 119:1<151 1996 119 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