title: Calcium-bindings of wild type and mutant troponin Cs of Caenorhabditis elegans.
abstract: Apparent Ca(2+)-binding constant (K(app)) of Caenorhabditis elegans troponin C (CeTnC) was determined by a fluorescence titration method. The K(app) of the N-domain Ca(2+)-binding site of CeTnC was 7.9+/-1.6 x 10(5) M(-1) and that of the C-domain site was 1.2+/-0.6 x 10(6) M(-1), respectively. Mg(2+)-dependence of the K(app) showed that both Ca(2+)-binding sites did not bind competitively Mg(2+). The Ca(2+) dissociation rate constant (k(off)) of CeTnC was determined by the fluorescence stopped-flow method. The k(off) of the N-domain Ca(2+)-binding site of CeTnC was 703+/-208 s(-1) and that of the C-domain site was 286+/-33 s(-1), respectively. From these values we could calculate the Ca(2+)-binding rate constant (k(on)) as to be 5.6+/-2.8 x 10(8) M(-1) s(-1) for the N-domain site and 3.4+/-2.1 x 10(8) M(-1) s(-1) for the C-domain site, respectively. These results mean that all Ca(2+)-binding sites of CeTnC are low affinity, fast dissociating and Ca(2+)-specific sites. Evolutional function of TnC between vertebrate and invertebrate and biological functions of wild type and mutant CeTnCs are discussed.
authors.affiliation: Department of Physics, School of Science, Nagoya University, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Ueda
authors.forename: H
authors.initials: H
authors.lastname: Katsuzaki
authors.forename: H
authors.initials: H
authors.lastname: Terami
authors.forename: H
authors.initials: H
authors.lastname: Ohtsuka
authors.forename: H
authors.initials: H
authors.lastname: Kagawa
authors.forename: T
authors.initials: T
authors.lastname: Murase
authors.forename: Y
authors.initials: Y
authors.lastname: Kajiwara
authors.forename: O
authors.initials: O
authors.lastname: Yoshioka
authors.forename: T
authors.initials: T
authors.lastname: Iio
mesh_terms: D000634:Aminoquinolines; D000818:Animals; D001665:Binding Sites; D017173:Caenorhabditis elegans / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D019143:Evolution, Molecular; D005456:Fluorescent Dyes; D007700:Kinetics; D009154:Mutation; D013050:Spectrometry, Fluorescence; D017186:Titrimetry; D019209:Troponin C / Q000235:genetics / Q000378:metabolism*
chemical_list: D000634:Aminoquinolines; D005456:Fluorescent Dyes; D019209:Troponin C; C035739:Quin2; D002118:Calcium