Organ distribution of rat histidine-pyruvate aminotransferase isoenzymes.
The organ distribution of rat histidine-pyruvate aminotransferase isoenzymes 1 and 2 was examined by using an isoelectric-focusing technique. Isoenzyme 1 (pI8.0) is present only in the liver and its activity is increased by the injection of glucagon, whereas isoenzyme 2 (pI5.2) is distributed in all tissues (liver, kidney, brain and heart) tested, and is not affected by glucagon injection. Isoenzyme 2 of the liver, kidney, brain and heart was purified by the same procedure and characterized. Isoenzyme 2 preparations from these four tissues were nearly identical in physical and enzymic properties. These properties differed from those previously found for the highly purified isoenzyme 1 preparation of rat liver. Isoenzyme 2 was active with pyruvate but not with 2-oxoglutarate as amino acceptor. Amino donors were effective in the following order of activity: tyrosine greater than histidine greater than phenylalanine greater than kynurenine greater than tryptophan. Very little activity was found with 5-hydroxytryptophan. The apparent Km for histidine was about 0.45 mM. The Km for pyruvate was about 4.5 mM with histidine as amino donor. The amino-transferase activities of isoenzyme 2 towards phenylalanine and tyrosine were inhibited by histidine. The ratio of aminotransferase activities towards these three amino acids was constant through gel filtration, electrophoresis, isoelectric focusing and sucrose-density-gradient centrifugation of the purified isoenzyme 2 preparations. These results suggest that these three activities are properties of the same enzyme protein. Sephadex G-150 gel filtration and sucrose-density-gradient centrifugation yielded mol.wts. of approx. 95000 and 92000 respectively. The pH optimum was between 9.0 and 9.3.
D000818:Animals; D001921:Brain / Q000201:enzymology; D005934:Glucagon / Q000494:pharmacology; D006639:Histidine; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000032:analysis*; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology; D009928:Organ Specificity; D011773:Pyruvates; D051381:Rats; D000637:Transaminases / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism
D007527:Isoenzymes; D011773:Pyruvates; D006639:Histidine; D005934:Glucagon; D000637:Transaminases