Factors affecting the amount and the activity of the glutamate dehydrogenases of Coprinus cinereus.
Kinetic analyses done with cell-free extracts of this basidiomycete fungus showed that the NADP-linked glutamate dehydrogenase exhibited positively co-operative interactions with the substrates 2-oxoglutarate and NADPH, negatively co-operative kinetics with NADP+ and was extremely sensitive to inhibition of deamination activity by ammonium and/or ammonia. The NAD-linked enzyme showed positive co-operativity with NADH, Michaelis-Menten kinetics with all other substrates and was subject only to mild inhibitions by the reaction products. Considered together with the values of the Michaelis constants, these results indicate that the former enzyme is primarily concerned with the amination of 2-oxoglutarate when the concentration of this substrate exceeds about 4 mM, while the NAD-linked enzyme is able to aminate or deaminate as metabolic conditions require. Synthesis of both enzymes was repressed by addition of carbamyl phosphate or N-acetyl-glutamate to mycelial cultures growing in media containing glucose and ammonium as carbon and nitrogen sources. Growth in media containing urea results in repression of the NADP-linked glutamate dehydrogenase and derepression of the NAD-linked enzyme. Such results indicate a connexion between the glutamate dehydrogenases and the urea cycle. It is suggested that under normal conditions of growth on complex media nitrogen is assimilated in the form of amino acids and that the glutamate dehydrogenases act in support of transaminases to allow this process to continue, and in support of the urea cycle to allow the disposal of excess nitrogen.
D000363:Agaricales / Q000201:enzymology*; D000494:Allosteric Regulation; D000641:Ammonia / Q000494:pharmacology; D003302:Coprinus / Q000201:enzymology*; D003470:Culture Media; D004789:Enzyme Activation; D004790:Enzyme Induction; D004794:Enzyme Repression; D005969:Glutamate Dehydrogenase / Q000378:metabolism*; D005971:Glutamates / Q000378:metabolism; D007656:Ketoglutaric Acids / Q000378:metabolism; D007700:Kinetics; D009243:NAD / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D014508:Urea / Q000378:metabolism
D003470:Culture Media; D005971:Glutamates; D007656:Ketoglutaric Acids; D009243:NAD; D009249:NADP; D000641:Ammonia; D014508:Urea; D005969:Glutamate Dehydrogenase