Kinetics of ATP hydrolysis by F1-ATPase and the effects of anion activation, removal of tightly bound nucleotides, and partial inhibition of the ATPase by covalent modification

Biochemistry ◽  
1984 ◽  
Vol 23 (21) ◽  
pp. 5004-5009 ◽  
Author(s):  
Siu Yin Wong ◽  
Akemi Matsuno-Yagi ◽  
Youssef Hatefi
Keyword(s):  
Atp Hydrolysis ◽  
Covalent Modification ◽  
F1 Atpase ◽  
Partial Inhibition ◽  
Kinetics Of ◽  
Tightly Bound Nucleotides

scholarly journals Kinetics of ATP hydrolysis by the F1-ATPase from Bacillus PS3: a reappraisal of the effects of ATP and Mg2+

1995 ◽  
Vol 1231 (1) ◽  
pp. 98-110 ◽  
Author(s):  
Stéphane Pezennec ◽  
Gérard Berger ◽  
Sandra Andrianambinintsoa ◽  
Nicolas Radziszewski ◽  
Guy Girault ◽  
...  
Keyword(s):  
Atp Hydrolysis ◽  
F1 Atpase ◽  
Kinetics Of

scholarly journals Bi-site activation occurs with the native and nucleotide-depleted mitochondrial F1-ATPase

1998 ◽  
Vol 330 (2) ◽  
pp. 1037-1043 ◽  
Author(s):  
M. Yakov MILGROM ◽  
B. Marat MURATALIEV ◽  
D. Paul BOYER
Keyword(s):  
Atp Hydrolysis ◽  
Maximal Activity ◽  
Catalytic Site ◽  
Turnover Rates ◽  
Velocity Measurements ◽  
F1 Atpase ◽  
Hydrolysis Rates ◽  
Recent Claim ◽  
Tightly Bound Nucleotides ◽  
Millimolar Concentration

Experiments are reported on the uni-site catalysis and the transition from uni-site to multi-site catalysis with bovine heart mitochondrial F1-ATPase. The very slow uni-site ATP hydrolysis is shown to occur without tightly bound nucleotides present and with or without Pi in the buffer. Measurements of the transition to higher rates and the amount of bound ATP committed to hydrolysis as the ATP concentration is increased at different fixed enzyme concentrations give evidence that the filling of a second site can initiate near maximal turnover rates. They provide rate constant information, and show that an apparent Km for a second site of about 2 μM and Vmax of 10 s-1, as suggested by others, is not operative. Careful initial velocity measurements also eliminate other suggested Km values and are consistent with bi-site activation to near maximal hydrolysis rates, with a Km of about 130 μM and Vmax of about 700 s-1. However, the results do not eliminate the possibility of additional ‘hidden’ Km values with similar Vmax:Km ratios. Recent data on competition between TNP-ATP and ATP revealed a third catalytic site for ATP in the millimolar concentration range. This result, and those reported in the present paper, allow the conclusion that the mitochondrial F1-ATPase can attain near maximal activity in bi-site catalysis. Our data also add to the evidence that a recent claim, that the mitochondrial F1-ATPase does not show catalytic site cooperativity, is invalid.

Transient kinetics of ATP hydrolysis by covalently crosslinked actomyosin complex in water and 40% ethylene glycol by the rapid flow quench method

Biochemistry ◽  
1985 ◽  
Vol 24 (14) ◽  
pp. 3814-3820 ◽  
Author(s):  
J. A. Biosca ◽  
F. Travers ◽  
T. E. Barman ◽  
R. Bertrand ◽  
E. Audemard ◽  
...  
Keyword(s):  
Ethylene Glycol ◽  
Atp Hydrolysis ◽  
Transient Kinetics ◽  
Actomyosin Complex ◽  
Rapid Flow ◽  
Kinetics Of ◽  
Quench Method

Molecular Mechanism of ATP Hydrolysis in F1-ATPase Revealed by Molecular Simulations and Single-Molecule Observations

2012 ◽  
Vol 134 (20) ◽  
pp. 8447-8454 ◽  
Author(s):  
Shigehiko Hayashi ◽  
Hiroshi Ueno ◽  
Abdul Rajjak Shaikh ◽  
Myco Umemura ◽  
Motoshi Kamiya ◽  
...  
Keyword(s):  
Molecular Mechanism ◽  
Single Molecule ◽  
Atp Hydrolysis ◽  
Molecular Simulations ◽  
F1 Atpase

scholarly journals Structure–property Effects in the Generation of Transient Aqueous Benzoic Acid Anhydrides by Carbodiimide Fuels

2019 ◽  
Author(s):  
Lasith Kariyawasam ◽  
Julie Kron ◽  
Run Jiang ◽  
André Sommer ◽  
Scott Hartley
Keyword(s):  
Atp Hydrolysis ◽  
Substituent Effects ◽  
Coupled Processes ◽  
Dissipative Systems ◽  
Nonequilibrium Systems ◽  
Structure Property ◽  
Ethylcarbodiimide Hydrochloride ◽  
Kinetics Of Formation ◽  
Acid Anhydrides ◽  
Kinetics Of

<div>The design of dissipative systems, which operate out-of-equilibrium by consuming chemical fuels, is challenging. As yet, there are few examples of privileged fuel chemistry that can be broadly applied in abiotic systems in the same way that ATP hydrolysis is exploited throughout biochemistry. The key issue is that designing nonequilibrium systems is inherently about balancing the relative rates of coupled processes. The use of carbodiimides as fuels to generate transient aqueous carboxylic anhydrides has recently been used in examples of new nonequilibrium materials and supramolecular assemblies. Here, we explore the kinetics of formation and decomposition of a series of benzoic anhydrides generated from the corresponding acids and EDC under prototypical conditions (EDC = <i>N</i>-(3-dimethylaminopropyl)-<i>N</i>′-ethylcarbodiimide hydrochloride). The reactions can be described by a simple mechanism that merges known behavior for the two processes independently. Structure–property effects in these systems are dominated by differences in anhydride decomposition rate. The kinetic parameters allow trends in concentration-dependent properties to be simulated, such as reaction lifetimes, peak anhydride concentrations, and efficiencies (i.e., total anhydride produced per equivalent of carbodiimide). For key properties there are diminishing returns with the addition of increasing amounts of fuel. This is particularly significant for the lifetimes, where substituent effects exert a much greater influence than fuel quantity under typical conditions. These results should provide useful guidelines for the design of functional systems making use of this chemistry.</div><div><br></div>

scholarly journals F1F0-ATP synthase from bovine heart mitochondria: development of the purification of a monodisperse oligomycin-sensitive ATPase

1993 ◽  
Vol 295 (3) ◽  
pp. 799-806 ◽  
Author(s):  
R Lutter ◽  
M Saraste ◽  
H S van Walraven ◽  
M J Runswick ◽  
M Finel ◽  
...  
Keyword(s):  
Atp Synthase ◽  
Atp Hydrolysis ◽  
Gel Filtration ◽  
Primary Objective ◽  
Membrane Domain ◽  
Mitochondrial Membranes ◽  
F1 Atpase ◽  
Ammonium Sulphate Precipitation ◽  
F1f0 Atp Synthase ◽  
Hydrolysis Activity

A new procedure for the isolation of ATP synthase from bovine mitochondria has been developed, with the primary objective of producing enzyme suitable for crystallization trials. Proteins were extracted from mitochondrial membranes with dodecyl-beta-D-maltoside, and the ATP synthase was purified from the extract in the presence of the same detergent by a combination of ion-exchange and gel-filtration chromatography and ammonium sulphate precipitation. This simple and rapid procedure yields 20-30 mg of highly pure and monodisperse enzyme, evidently consisting of 14 different subunits, amongst them, in apparently stoichiometric amounts with the established subunits, subunit e, a recently discovered subunit of unknown function. The enzyme preparation has an oligomycin-sensitive ATP hydrolysis activity, and so the F1 domain is functionally associated with the membrane domain, F0. In contrast with the N-termini of some of the subunits of bovine mitochondrial F1-ATPase, those of the F1F0-ATP synthase are not degraded by proteolysis during the isolation procedure. This preparation therefore satisfies prerequisites for crystallization trials.

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