Biomaterial Database

Mitochondrial adenosine triphosphatase from yeast, Saccharomyces cerevisiae. Purification, subunit structure and kinetics. 1976

K Takeshige, and B Hess, and M Böhm, and H Zimmermann-Telschow

1. A procedure for the purification of ATPase extracted by chloroform from baker's yeast (Saccharomyces cerevisiae) is reported. The yield based on submitochondrial particles was 55% and the purification was 100-fold. The isolated complex was homogenous as assessed by gel filtration, ion-exchange chromatography, sedimentation in sucrose gradient and in the analytical ultracentrifuge. The molecular weight determined by gel filtration was 400000 +/- 20000. Ultracentrifugation yielded s020,w = 12.50 +/- 0.13 S and the laser light scattering study gave a diffusion coeficient of D20w - 2.92 X 10(-7) cm2 s-1. The amino acid composition as well as absorption, fluorescence, and circular dichroism spectra, from which the helicity of 39% was evaluated, are given. 2. On polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate, six components with molecular weights of 58500(alpha), 55000 (beta), 42000, 34000 (gamma), 10000(delta), and 8600 (epsilon) were observed with a stoichiometry of 3:3:1:1:1:1. The amino acid composition is given for alpha + beta and gamma as well as delta and epsilon components. 3. The maximum specific activity of the enzyme was 200 U/mg under the optimum conditions. The enzyme was inactivated by incubation at 0 degrees C and strongly inhibited by the antibiotic Dio-9 but not by oligomycin and N, N'-dicyclohexyl-carbodiimide. The effects of kinetic parameters and anions on the enzyme are reported. Two active sites for Mg-ATP with Km values of 0.045mM and 0.37mM and a single activie site for Mg-ITP with Km = 0.179mM were found. A study of the temperature dependence of the maximum activity revealed a straight line in the Arrhenius plots with an activation energy of 11.0 kcal/mol (= 46 kH/mol).

UI	MeSH Term	Description	Entries
D007700	Kinetics	The rate dynamics in chemical or physical systems.
D008928	Mitochondria	Semiautonomous, self-reproducing organelles that occur in the cytoplasm of all cells of most, but not all, eukaryotes. Each mitochondrion is surrounded by a double limiting membrane. The inner membrane is highly invaginated, and its projections are called cristae. Mitochondria are the sites of the reactions of oxidative phosphorylation, which result in the formation of ATP. They contain distinctive RIBOSOMES, transfer RNAs (RNA, TRANSFER); AMINO ACYL T RNA SYNTHETASES; and elongation and termination factors. Mitochondria depend upon genes within the nucleus of the cells in which they reside for many essential messenger RNAs (RNA, MESSENGER). Mitochondria are believed to have arisen from aerobic bacteria that established a symbiotic relationship with primitive protoeukaryotes. (King & Stansfield, A Dictionary of Genetics, 4th ed)	Mitochondrial Contraction,Mitochondrion,Contraction, Mitochondrial,Contractions, Mitochondrial,Mitochondrial Contractions
D009840	Oligomycins	A closely related group of toxic substances elaborated by various strains of Streptomyces. They are 26-membered macrolides with lactone moieties and double bonds and inhibit various ATPases, causing uncoupling of phosphorylation from mitochondrial respiration. Used as tools in cytochemistry. Some specific oligomycins are RUTAMYCIN, peliomycin, and botrycidin (formerly venturicidin X).	Oligomycin
D000251	Adenosine Triphosphatases	A group of enzymes which catalyze the hydrolysis of ATP. The hydrolysis reaction is usually coupled with another function such as transporting Ca(2+) across a membrane. These enzymes may be dependent on Ca(2+), Mg(2+), anions, H+, or DNA.	ATPases,Adenosinetriphosphatase,ATPase,ATPase, DNA-Dependent,Adenosine Triphosphatase,DNA-Dependent ATPase,DNA-Dependent Adenosinetriphosphatases,ATPase, DNA Dependent,Adenosinetriphosphatases, DNA-Dependent,DNA Dependent ATPase,DNA Dependent Adenosinetriphosphatases,Triphosphatase, Adenosine
D000596	Amino Acids	Organic compounds that generally contain an amino (-NH2) and a carboxyl (-COOH) group. Twenty alpha-amino acids are the subunits which are polymerized to form proteins.	Amino Acid,Acid, Amino,Acids, Amino
D000838	Anions	Negatively charged atoms, radicals or groups of atoms which travel to the anode or positive pole during electrolysis.	Anion
D012441	Saccharomyces cerevisiae	A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement.	Baker's Yeast,Brewer's Yeast,Candida robusta,S. cerevisiae,Saccharomyces capensis,Saccharomyces italicus,Saccharomyces oviformis,Saccharomyces uvarum var. melibiosus,Yeast, Baker's,Yeast, Brewer's,Baker Yeast,S cerevisiae,Baker's Yeasts,Yeast, Baker
D013696	Temperature	The property of objects that determines the direction of heat flow when they are placed in direct thermal contact. The temperature is the energy of microscopic motions (vibrational and translational) of the particles of atoms.	Temperatures
D046911	Macromolecular Substances	Compounds and molecular complexes that consist of very large numbers of atoms and are generally over 500 kDa in size. In biological systems macromolecular substances usually can be visualized using ELECTRON MICROSCOPY and are distinguished from ORGANELLES by the lack of a membrane structure.	Macromolecular Complexes,Macromolecular Compounds,Macromolecular Compounds and Complexes,Complexes, Macromolecular,Compounds, Macromolecular,Substances, Macromolecular