Activity of lactate dehydrogenase isoenzymes LD1 and LD2 in serum as determined by using an inhibitor of the M-subunit.
To measure activities of lactate dehydrogenase (EC 1.1.1.27) isoenzymes LD1 and LD2 in serum, we developed a method involving 1,6-hexanediol as specific inhibitor of the M-subunit. Addition of hexanediol, 0.6 mol/L, to five LD isoenzyme fractions purified from human liver and heart homogenates resulted in complete loss of activities of LD4 and LD5, and partial loss of LD2 and LD3. The activity of LD1, which is composed of the H-subunit only, was not affected. In studying what conditions would allow only the activities of LD1 or LD1 + LD2 to be expressed in serum, we found that the respective activities could be determined by treatment with hexanediol, 0.75 mol/L and 0.55 mol/L, respectively. Results of binding experiments and analytical-recovery tests supported the effectiveness of analyses with this inhibitor in determination of LD1 and LD1 + LD2 activities in serum. Results by this proposed inhibition method correlated well with those by the conventional electrophoretic method for determination of LD1 activity, but LD1 + LD2 activities by the inhibition method were a little less than those by the electrophoretic method, requiring some correction.
D004591:Electrophoresis, Polyacrylamide Gel; D006018:Glycols / Q000494:pharmacology; D006331:Heart Diseases / Q000201:enzymology; D006801:Humans; D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase / Q000037:antagonists & inhibitors / Q000097:blood*; D008107:Liver Diseases / Q000201:enzymology; D046911:Macromolecular Substances; D013997:Time Factors
D006018:Glycols; D007527:Isoenzymes; D046911:Macromolecular Substances; D007770:L-Lactate Dehydrogenase; C027765:hexamethylene glycol