Nitrogen-13 flux from L-[13N]glutamate in the isolated rabbit heart: effect of substrates and transaminase inhibition.
Kinetic and biochemical parameters of nitrogen-13 flux from L-[13N]glutamate in myocardium were examined. Tissue radioactivity kinetics and chemical analyses were determined after bolus injection of L-[13N]glutamate into isolated arterially perfused interventricular septa under various metabolic states, which included addition of lactate, pyruvate, aminooxyacetate (a transaminase inhibitor), or a combination of aminooxyacetate and pyruvate to the standard perfusate containing insulin and glucose. Chemical analysis of tissue and effluent at 6 min allowed determination of the composition of the slow third kinetic component of the time-activity curves. 13N-labeled aspartate, alanine and glutamate accounted for more than 80% of the tissue nitrogen-13 under the experimental conditions used. Specific activities for these amino acids were constant, but not identical to each other, from 6 through 15 min after administration of L-[13N]glutamate. Little labeled ammonia (1.9%) and glutamine (4.7%) were produced, indicating limited accessibility of exogenous glutamate to catabolic mitochondrial glutamate dehydrogenase and glutamine synthetase, under control conditions. Lactate and pyruvate additions did not affect tissue amino acid specific activities. Aminooxyacetate suppressed formation of 13N-labeled alanine and aspartate and increased production of L-[13N]glutamine and [13N]ammonia. Formation of [13N]ammonia was, however, substantially decreased when aminooxyacetate was used in the presence of exogenous pyruvate. The data support a model for glutamate compartmentation in myocardium not affected by increasing the velocity of enzymatic reactions through increased substrate (i.e., lactate or pyruvate) concentrations but which can be altered by competitive inhibition of transaminases (via aminooxyacetate) making exogenous glutamate more available to other compartments.
D000085:Acetates / Q000494:pharmacology*; D000410:Alanine Transaminase / Q000037:antagonists & inhibitors*; D000625:Aminooxyacetic Acid / Q000494:pharmacology*; D000641:Ammonia / Q000378:metabolism; D000818:Animals; D001219:Aspartate Aminotransferases / Q000037:antagonists & inhibitors*; D005971:Glutamates / Q000378:metabolism*; D005973:Glutamine / Q000378:metabolism; D066298:In Vitro Techniques; D007700:Kinetics; D007773:Lactates / Q000494:pharmacology*; D009206:Myocardium / Q000378:metabolism*; D009590:Nitrogen Radioisotopes; D011773:Pyruvates / Q000494:pharmacology*; D011817:Rabbits
D000085:Acetates; D005971:Glutamates; D007773:Lactates; D009590:Nitrogen Radioisotopes; D011773:Pyruvates; D005973:Glutamine; D000625:Aminooxyacetic Acid; D000641:Ammonia; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase