Effect of acute metabolic acidosis on ammonia metabolism in kidney.
To understand the mechanisms that initiate the increase in ammonia formation during acute acidosis in kidney [amino-15N]- and [amino-15N]glutamine were used as substrates in isolated perfused rat kidney experiments. Perfused kidneys from methionine sulfoximine-treated rats take up glutamine nitrogen at the rate of 1.50 +/- 0.08 mumol.g kidney-1.min-1 while forming ammonia at a rate of 0.65 +/- 0.09 mumol.g.kidney-1.min-1. Mass spectrometer analysis of the perfusate and urine reveals that ammonia is formed from the amide nitrogen of glutamine at the rate of 0.32 +/- 0.06 mumol.g kidney-1.min-1 and ammonia is formed from glutamate derived from glutamine at the rate of 0.21 +/- 0.04 mumol.g kidney-1.min-1. The balance of the ammonia formed is from unidentified endogenous sources. Addition of HCl to the perfusate to lower perfusate pH increases ammonia formation to 1.09 +/- 0.10 mumol.g kidney-1.min-1. The results exclude a role for the purine nucleotide cycle during acute acidosis and confirm that ammonia formation from glutamate derived from glutamine is via glutamate dehydrogenase. Lowering perfusate pH increases the rate of glutamine deamidation significantly by 0.33 +/- 0.06 mumol.g kidney-1.min-1 and increases the rate of ammonia formation via glutamate dehydrogenase insignificantly by only 0.08 +/- 0.04 mumol.g kidney-1.min-1, whereas ammonia formation from endogenous sources remains unchanged. The results demonstrate that regulation of glutamine deamidation is an important controlling step in ammonia formation during acute metabolic acidosis in kidney.
Bogusky
R T
RT
Department of Internal Medicine, School of Medicine, University of California, Davis, Sacramento 95616.
Dietrich
R L
RL
D000138:Acidosis / Q000378:metabolism*; D000208:Acute Disease; D000227:Adenine Nucleotides / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism; D000641:Ammonia / Q000378:metabolism*; D000818:Animals; D008401:Gas Chromatography-Mass Spectrometry; D005973:Glutamine / Q000378:metabolism; D006150:Guanine Nucleotides / Q000378:metabolism; D066298:In Vitro Techniques; D007291:Inosine Monophosphate / Q000378:metabolism; D007668:Kidney / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D009587:Nitrogen Isotopes; D010477:Perfusion; D051381:Rats; D011919:Rats, Inbred Strains; D012016:Reference Values
D000227:Adenine Nucleotides; D000596:Amino Acids; D006150:Guanine Nucleotides; D009587:Nitrogen Isotopes; D005973:Glutamine; D007291:Inosine Monophosphate; D000641:Ammonia