Activity of heart and muscle lactate dehydrogenases in all-aqueous systems and in organic solvents with low amounts of water. Effect of guanidine chloride.
The effect of urea and guanidine hydrochloride (GdmCl) on the activity of lactate dehydrogenases from heart and muscle was studied in standard water mixtures and in reverse micelles formed with n-octane, hexanol, cetyltrimethylammonium bromide and water in a concentration that ranged over 2.5-6.0% (by vol.). In all water mixtures GdmCl (0.15-0.75 M) and urea (0.5-3.0 M) inhibited the activity of the enzymes at non-saturating pyruvate concentrations. At concentrations of pyruvate that proved inhibitory for enzyme activity due to the formation of a ternary enzyme-NAD-pyruvate complex, GdmCl and urea increased the activity of the enzymes. This increase correlated with a decrease of the ternary complex, as evidenced by its absorbance at 320-325 nm. In the low-water system it was found that: (a) at all concentrations of pyruvate tested (0.74-30 mM), GdmCl enhanced the activity of the heart enzyme to a similar extent; (b) in the muscle enzyme, GdmCl inhibited or increased the activity through a process that depended on the concentration of pyruvate and GdmCl; (c) under optimal conditions, the activation by GdmCl was about two times lower in the muscle than in the heart enzyme, although in all-water media the activity of the muscle enzyme was twice as high. The expression of lactate dehydrogenase activity in the low-water system was higher with the heart than with the muscle enzyme compared to their activities in all-water media (about 260 and 600 mumol min-1 mg-1 in the heart and muscle enzymes respectively). Apparently for catalysis, the water requirement in the heart enzyme is lower than in the muscle enzyme. It is likely that the different response of the two enzymes to solvent is due to their distinct structural features.
Fernández-Velasco
D A
DA
Instituto de Fisiología Celular, Universidad Nacional Autónoma de México.
Garza-Ramos
G
G
Ramírez
L
L
Shoshani
L
L
Darszon
A
A
Tuena de Gómez-Puyou
M
M
Gómez-Puyou
A
A
D000818:Animals; D002417:Cattle; D019791:Guanidine; D006146:Guanidines / Q000494:pharmacology*; D007527:Isoenzymes; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism*; D008823:Micelles; D009132:Muscles / Q000201:enzymology*; D009206:Myocardium / Q000201:enzymology*; D012997:Solvents; D013056:Spectrophotometry, Ultraviolet; D013552:Swine; D014508:Urea / Q000494:pharmacology; D014867:Water
D006146:Guanidines; D007527:Isoenzymes; D008823:Micelles; D012997:Solvents; D014867:Water; D014508:Urea; D007770:L-Lactate Dehydrogenase; D019791:Guanidine