Isolation and characterisation of tissue-specific isoenzymes of D-lactate dehydrogenase from muscle and hepatopancreas ofLimulus polyphemus
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Isoenzymes of dimeric,d-lactate specific lactate dehydrogenase from hepatopancreas and muscle tissue of the horseshoe crab,Limulus polyphemus, have been purified to homogeneity by chromatographic techniques.
Separation of the three muscle isoenzymes, designated CC, CD and DD, was achieved by ion-exchange chromatography, whereas the three hepatopancreas isoenzymes, designated AA, AB and BB, were separated by chromatofocusing.
Kinetic studies suggest that hepatopancreasd-LDH is analogous to vertebrate heart-typel-LDH; whereas horseshoe crab muscled-LDH resembles vertebrate M-typel-LDH; for example, hepatopancreasd-LDH is strongly substrate inhibited by pyruvate, whereas muscled-LDH is not. Moreover, hepatopancreasd-LDH has a higher affinity ford-lactate and NAD+ than muscled-LDH. Finally, at physiological pH values, it appears that the hepatopancreas isoenzymes are strongly poised in the direction ofd-lactate oxidation.
The kinetic data indicate the potential for organ partitioning ofd-lactate formation andd-lactate oxidation:d-lactate produced in the muscles may be transported via the hemolymph to the hepatopancreas for oxidation.
KeywordsLactate Pyruvate Human Physiology Muscle Tissue Lactate Dehydrogenase
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