Cellular and subcellular localization of enzymes of arginine metabolism in rat kidney.
Rat kidneys extract citrulline derived from the intestinal metabolism of glutamine and convert it stoichiometrically into arginine. This pathway constitutes the major endogenous source of arginine. We investigated the localization of enzymes of arginine synthesis, argininosuccinate synthase and lyase, and of breakdown, arginase and ornithine aminotransferase, in five regions of rat kidney, in cortical tubule fractions and in subcellular fractions of cortex. Argininosuccinate synthase and lyase were found almost exclusively in cortex. Arginase and ornithine aminotransferase were found in inner cortex and outer medulla. Since cortical tissue primarily consists of proximal convoluted and straight tubules, distal tubules and glomeruli, we prepared cortical tubule fragments by collagenase digestion of cortices and fractionated them on a Percoll gradient. Argininosuccinate synthase and lyase were found to be markedly enriched in proximal convoluted tubules, whereas less than 10% of arginase and ornithine aminotransferase, were recovered in this fraction. Arginine production from citrulline was also enriched in proximal convoluted tubules. Subcellular fractionation of kidney cortex revealed that argininosuccinate synthase and lyase are cytosolic. We therefore conclude that arginine synthesis occurs in the cytoplasm of the cells of the proximal convoluted tubule.
Dhanakoti
S N
SN
Department of Biochemistry, Memorial University of Newfoundland, St. John's, Canada.
Brosnan
M E
ME
Herzberg
G R
GR
Brosnan
J T
JT
D000818:Animals; D001119:Arginase / Q000378:metabolism; D001120:Arginine / Q000378:metabolism*; D001123:Argininosuccinate Lyase / Q000378:metabolism*; D001124:Argininosuccinate Synthase / Q000378:metabolism*; D005966:Glucuronidase / Q000378:metabolism; D006593:Hexokinase / Q000378:metabolism; D007672:Kidney Cortex / Q000201:enzymology*; D007679:Kidney Medulla / Q000201:enzymology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism; D009953:Ornithine-Oxo-Acid Transaminase / Q000378:metabolism; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013384:Succinate Cytochrome c Oxidoreductase / Q000378:metabolism
D001120:Arginine; D013384:Succinate Cytochrome c Oxidoreductase; D007770:L-Lactate Dehydrogenase; D009251:NADPH-Ferrihemoprotein Reductase; D009953:Ornithine-Oxo-Acid Transaminase; D006593:Hexokinase; D005966:Glucuronidase; D001119:Arginase; D010729:Phosphoenolpyruvate Carboxykinase (GTP); D001123:Argininosuccinate Lyase; D001124:Argininosuccinate Synthase