Kinetics of slow reversible inhibition of human muscle creatine kinase by planar anions.
The toxicity of NO3- and NO2- to mammals has been widely publicized. However, the kinetic mechanism of inhibition of human muscle creatine kinase by NO3- and NO2- has not been explored. The kinetic theory of the substrate reaction during the modification of enzyme activity previously described by Tsou (Adv. Enzymol. Related Areas Mol. Biol. 1988, 61, 381-436) has been applied to a study of the kinetics of slow reversible inhibition of human muscle creatine kinase by planar anions (NO3- and NO2-). The kinetic equation of the substrate reaction was derived from theoretical analysis and experimental data, then simplified. The microscopic rate constants for the reaction of the inhibitors with the enzyme were obtained from the simplified equation for the substrate reaction in the presence of the inhibitors. The results show that the apparent forward rate constant A is dependent on ATP concentration, indicating competition between the inhibitor (NO3- or NO2-) and ATP. The results also suggest that binding of creatine-MgADP and the anion with the enzyme is very tight, since their binding constants are much higher than those for normal substrates.
Department of Biological Science and Biotechnology, Tsinghua University, Beijing, 100084, China.
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Luo
W Z
WZ
Xie
J H
JH
Bai
H M
HM
Zhou
D000255:Adenosine Triphosphate / Q000378:metabolism; D001667:Binding, Competitive; D003402:Creatine Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes; D007700:Kinetics; D008956:Models, Chemical; D018482:Muscle, Skeletal / Q000201:enzymology*; D009566:Nitrates / Q000494:pharmacology; D009573:Nitrites / Q000494:pharmacology
D007527:Isoenzymes; D009566:Nitrates; D009573:Nitrites; D000255:Adenosine Triphosphate; D003402:Creatine Kinase