Kinetic behaviour of succinate dehydrogenase of three fibre types in skeletal muscle. I. Effects of temperature and a competitive inhibitor.
The kinetic behaviour of succinate dehydrogenase [EC 1.3.99.1] in three fibre types of rat gastrocnemius was examined by a quantitative histochemical method without disruption of the cellular structure. 2-(2-Benzothiazolyl)-3-(4-phthalhydrazidyl)-5-styryl-t etrazolium chloride (BPST) and phenazine methosulphate were used as electron acceptors. On measurement of the absorbance value at 530 nm of BPST formazan, produced by the succinate dehydrogenase reaction in sections, it was found that the staining intensity of succinate dehydrogenase was linearly proportional to both the incubation time and the thickness of the slice therefore, the initial velocity of the staining could be calculated. By Michaelis-Menten (1913) treatment of the dependence of the initial velocity on the substrate concentration in the absence and the presence of a competitive inhibitor, malonate, the Km and Vmax values for succinate and the Ki value for malonate were obtained. The Km and Ki values of the three fibre types were similar. The ration of the Vmax values of type A, B and C fibres was 1.0:2.0:3.3. The temperature dependence of the kinetic parameters was very similar in the three fibre types. These findings confirm that the differences in the staining intensity of the three fibre types reflect differences in the amounts, but not the properties, of succinate dehydrogenase.
D000818:Animals; D001667:Binding, Competitive; D007700:Kinetics; D008297:Male; D009132:Muscles / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains; D013194:Staining and Labeling; D013385:Succinate Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013696:Temperature
D013385:Succinate Dehydrogenase