Deviations from Michaelis-Menten kinetics. The possibility of complicated curves for simple kinetic schemes and the computer fitting of experimental data for acetylcholinesterase, acid phosphatase, adenosine deaminase, arylsulphatase, benzylamine oxidase, chymotrypsin, fumarase, galactose dehydrogenase, beta-galactosidase, lactate dehydrogenase, peroxidase and xanthine oxidase.
The possible graph shapes for one-site/two-state and substrate-modifier models are discussed. The two-state model is a version of the Monod-Wyman-Changeux model and gives a rate equation with 240 denominator terms. Discussion in terms of K and V effects is not possible. A simplified version of the mechanism can be shown to give v-versus-[S] curves that are either sigmoid or non-sigmoid. They may show substrate inhibition or no final maximum, and the double-reciprocal plots can be concave up or down. The corresponding binding model is determined by only two constants and gives a linear double-reciprocal plot. The substrate-modifier mechanism is a simple example of a mechanism where inclusion of catalytic steps leads to a genuine increase in degree of the rate equation. The v-versus-[S] curve can show such complexities as two maxima and a minimum, and the double-reciprocal plot can cross its asymptote twice, proving the rate equation to be 4:4. A simplified version is 3:3, and analysis shows that at least 18 of the 27 double-reciprocal plots that can arise with 3:3 functions are possible with this particular mechanism. Representative double-reciprocal and Scatchard plots are presented for several sets of rate-constant values. It is concluded that relatively simple mechanisms give pseudo-steady-state rate equations of high degree and considerable complexity. With extended ranges of substrate concentrations there is every reason to believe that experimental data would show the sort of deviations from Michaelis-Menten kinetics seen with calculated curves for such simple mechanisms. Narrow ranges of substrate concentration, on the other hand, would lead to inflexions and curvature being overlooked. It is not possible to discuss such deviations from Michaelis-Menten kinetics in terms of kinetic constants such as Km and V, and, in general, it is also difficult to see any simple way to explain intuitively such features as sigmoidicity, substrate inhibition, double-reciprocal convexity and decrease in degree by cancellation of common factors between numerator and denominator of rate equations. These conclusions apply with even more force when catalytic steps are included, for then the rate equations, are for multi-site mechanisms, of higher degree, allowing increasingly complex curve shapes. A number of enzymes were studied and initial-rate data were fitted by computer.(ABSTRACT TRUNCATED AT 400 WORDS)
D000110:Acetylcholinesterase / Q000378:metabolism; D000135:Acid Phosphatase / Q000378:metabolism; D000243:Adenosine Deaminase / Q000378:metabolism; D001192:Arylsulfatases / Q000378:metabolism; D001595:Benzylamine Oxidase / Q000378:metabolism; D002918:Chymotrypsin / Q000378:metabolism; D003201:Computers; D005649:Fumarate Hydratase / Q000378:metabolism; D005691:Galactose Dehydrogenases / Q000378:metabolism; D006867:Hydrolases / Q000378:metabolism*; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008433:Mathematics; D010088:Oxidoreductases / Q000378:metabolism*; D010544:Peroxidases / Q000378:metabolism; D014969:Xanthine Oxidase / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
D010088:Oxidoreductases; D005691:Galactose Dehydrogenases; D007770:L-Lactate Dehydrogenase; D010544:Peroxidases; D014969:Xanthine Oxidase; D001595:Benzylamine Oxidase; D006867:Hydrolases; D000110:Acetylcholinesterase; D000135:Acid Phosphatase; D001192:Arylsulfatases; D001616:beta-Galactosidase; D002918:Chymotrypsin; D000243:Adenosine Deaminase; D005649:Fumarate Hydratase