Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.6.3.14 (ATP synthase)
 7,042 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Bovine heart mitochondrial ATPase is inhibited after covalent modification with 4-chloro-7-nitrobenzofuroxan. The kinetics of the reaction are indistinguishable from those for the reaction of an essential tyrosine residue of the ATPase with 4-chloro-7-nitrobenzofurazan that have been described previously [Ferguson et al. (1975) Eur. J. Biochem. 54, 117-126]. 4-Fluoro-7-nitrobenzofurazan inhibits the ATPase with a pseudo-first-order rate constant that is tenfold greater than that for 4-chloro-7-nitrobenzofurazan. These data indicate that the rate-limiting step for reaction of the enzyme with these reagents is formation of a Meisenheimer complex at the C-4 position and that the modified tyrosine is probably on the surface of the protein. No evidence was found for more complex patterns of reactivity of 4-chloro-7-nitrobenzofurazan and its analogues. Both ammonium 4-chloro-7-sulphobenzofurazan and ammonium 4-fluoro-7-sulphobenzofurazan fail to react with the ATPase. The utility of these reagents as alternatives to the nitro derivatives may be limited owing to their slow reaction rates. After modification on tyrosine by 4-chloro-7-nitrobenzofurazan, the nitrobenzofurazan group can be transferred by an intramolecular process to lysine [Ferguson et al. (1975) Eur. J. Biochem. 54, 127-133]. ATPase with the lysine thus modified is shown to be reactive towards 4-chloro-7-nitrobenzofurazan in a manner indistinguishable from the native enzyme. This indicates that the intramolecular transfer occurs at sufficient distance to avoid steric hindrance to the second reaction, and that the lysine does not participate in a neighbouring group effect to enhance the reactivity of the tyrosine.
 ...
PMID:The nature of the reaction of an essential tyrosine residue of bovine heart mitochondrial ATPase with 4-chloro-7-nitrobenzofurazan and related compounds. 623 12

1. The ATPase activity of insect mitochondria has been investigated. A comparison was made to determine the distribution and nature of such activity in other isolated fractions of the house fly, Musca domestica L. 2. The ATPase in insect mitochondria is specific in that orthophosphate can be cleaved only from ATP. The Michaelis-Menten constant K(8) = 2.78 x 10(-3)M and V(max.) = 76 micrograms P min.(-1) mg.(-1) dry weight. 3. Mg(++) and Mn(++) activate this enzymatic reaction in mitochondria, but Ca(++) does not. The extent of activation is 60 per cent with the optimal concentration 6 x 10(-4)M. Experiments with combinations of Mg(++) and Mn(++) show that either ion can replace the other and that the effects are additive, depending solely on the final concentration of the combination. Concentrations of Mg, Mn, or Ca ions higher than 6 x 10(-3)M inhibit the enzyme. 4. Fluoride does not inhibit the ATPase of insect mitochondria, whereas azide and chloromercuribenzoate do. The per cent inhibition depends on the concentration of inhibitor. 5. Finely dispersed mitochondrial particles have much greater ATPase activity than intact mitochondria. The possible relationship of this observation to latent ATPase is considered. 6. A magnesium-activated adenylate kinase is present in these mitochondria. The liberated orthophosphate, derived from ADP, is the result of the activity of adenylate kinase followed by the specific ATPase. 7. ATP can be dephosphorylated by enzymes found in the muscle fibrils, and in a "soluble" fraction, as well as in mitochondria. The fibrillar ATPase is Ca(++)-activated. The "soluble" fraction, however, like the mitochondria, is Mg(++)-activated. The "soluble" ATP dephosphorylation mechanism is distinguished from the mitochondrial ATPase in that it is inhibited by fluoride. 8. The "soluble" fraction also contains a magnesium-activated inorganic pyrophosphatase. Fluoride completely inhibits this enzymatic reaction. 9. The possible mechanism of ATP dephosphorylation in the "soluble" fraction is discussed.
 ...
PMID:Investigations on the mitochondria of the house fly, Musca domestica L. I. Adenosinetriphosphatases. 1302 33

F(0)F(1)-ATP synthases couple proton translocation with the synthesis of ATP from ADP and phosphate. The enzyme has three catalytic nucleotide binding sites, one on each beta-subunit; three non-catalytic binding sites are located mainly on each alpha-subunit. In order to observe substrate binding to the enzyme, the H(+)-ATP synthase from Escherichia coli was labelled selectively with the fluorescence donor tetramethylrhodamine (TMR) at position T106C of the gamma-subunit. The labelled enzymes were incorporated into liposomes and catalysed proton-driven ATP synthesis. The substrate ATP-Alexa Fluor 647 was used as the fluorescence acceptor to perform intermolecular fluorescence resonance energy transfer (FRET). Single molecules are detected with a confocal set-up. When one ATP-Alexa Fluor 647 binds to the enzyme, FRET can be observed. Five stable states with different intermolecular FRET efficiencies were distinguished for enzyme-bound ATP-Alexa Fluor 647 indicating binding to different binding sites. Consecutive hydrolysis of excess ATP resulted in stepwise changes of the FRET efficiency. Thereby, gamma-subunit movement during catalysis was directly monitored with respect to the binding site with bound ATP-Alexa Fluor 647.
 ...
PMID:Binding of single nucleotides to H+-ATP synthases observed by fluorescence resonance energy transfer. 1511 Feb 52