title: Functions and ATP-binding responses of the twelve histidine residues in the TF1-ATPase beta subunit.
abstract: The C2 proton signals of all (twelve) histidine residues of the TF1 beta subunit in the 1H-NMR spectrum have been identified and assigned by means of pH change experiments and site-directed substitution of histidines by glutamines. pH and ligand titration experiments were carried out for these signals. Furthermore, the ATPase activity of the reconstituted alpha3beta3gamma complex was examined for the twelve mutant beta subunits. Two of three conserved histidines, namely, His-119 and 324, were found to be important for expression of the ATPase activity. The former fixes the N-terminal domain to the central domain. His-324 is involved in the formation of the interface essential for the alpha3beta3gamma complex assembly. The other conserved residue, His-363, showed a very low pK(a), suggesting that it is involved in the tertiary structure formation. On the binding of a nucleotide, only the signals of His-173, 179, 200, and 324 shifted. These histidines are located in the hinge region, and its proximity, of the beta subunit. This observation provided further support for the conformational change of the beta monomer from the open to the closed form on the binding of a nucleotide proposed by us [Yagi et al. (1999) Biophys. J. 77, 2175-2183]. This conformational change should be one of the essential driving forces in the rotation of the alpha3beta3gamma complex.
authors.affiliation: Department of Chemistry and Biotechnology, Faculty of Engineering, Yokohama National University, Hodogaya-ku, Yokohama 240-8501, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Tozawa
authors.forename: H
authors.initials: H
authors.lastname: Yagi
authors.forename: K
authors.initials: K
authors.lastname: Hisamatsu
authors.forename: K
authors.initials: K
authors.lastname: Ozawa
authors.forename: M
authors.initials: M
authors.lastname: Yoshida
authors.forename: H
authors.initials: H
authors.lastname: Akutsu
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D019943:Amino Acid Substitution; D025243:Bacterial Proton-Translocating ATPases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D020134:Catalytic Domain; D017124:Conserved Sequence; D004926:Escherichia coli / Q000235:genetics; D006639:Histidine / Q000235:genetics / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles / Q000737:chemistry; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D019906:Nuclear Magnetic Resonance, Biomolecular; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D021122:Protein Subunits; D011522:Protons
chemical_list: D007093:Imidazoles; D021122:Protein Subunits; D011522:Protons; D006639:Histidine; D000255:Adenosine Triphosphate; D025243:Bacterial Proton-Translocating ATPases