N omega-phosphoarginine phosphatase from rat renal microsome was alkaline phosphatase.
Activity hydrolyzing both N omega-phosphoarginine and glucose-6-phosphate was detected in rat renal microsome but not in hepatic microsome. Renal microsome was solubilized with 1% n-octyl-beta-D-thioglucoside and purified with DEAE-Sepharose column chromatography. Fractions hydrolyzing both N omega-phosphoarginine or glucose-6-phosphate were subjected to 7.5%-polyacrylamide gel electrophoresis in the presence of 0.1% sodium dodecyl sulfate. Phosphatase activity in the gels was detected by a lead nitrate stain using N omega-phosphoarginine or glucose-6-phosphate as substrates. Both substrates produced a stain in the region of the gel corresponding to a protein with a mass of 150 kDa. Extracts of slices from this region of the gel also hydrolyzed phosphocreatine, inorganic pyrophosphate, and O-phosphotyrosine. Moreover, the phosphatase had its optimal pH in the alkaline range and was inhibited completely by 20 microM sodium vanadate, 1 mM cysteine, and 1 mM tetramisole. All these properties indicate that the microsomal phosphoamidase (EC 3.9.1.1) of rat kidney was identical with alkaline phosphatase (EC 3.1.3.1).
Department of Biochemistry, Saga Medical School, Saga-ken, Japan.
M
M
Nishino
S
S
Tsujimura
M
M
Kuba
A
A
Kumon
D000469:Alkaline Phosphatase / Q000378:metabolism*; D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives* / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology; D004795:Enzyme Stability; D005952:Glucose-6-Phosphatase / Q000187:drug effects / Q000378:metabolism; D019298:Glucose-6-Phosphate; D005958:Glucosephosphates / Q000378:metabolism*; D006867:Hydrolases / Q000187:drug effects / Q000378:metabolism; D007668:Kidney / Q000201:enzymology*; D008297:Male; D008670:Metals / Q000494:pharmacology; D008861:Microsomes / Q000201:enzymology*; D008862:Microsomes, Liver / Q000201:enzymology; D009943:Organophosphorus Compounds / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000302:isolation & purification; D051381:Rats; D017208:Rats, Wistar; D013379:Substrate Specificity; D013438:Sulfhydryl Compounds / Q000494:pharmacology; D013644:Tartrates / Q000494:pharmacology; D013773:Tetramisole / Q000494:pharmacology
D003902:Detergents; D005958:Glucosephosphates; D008670:Metals; D009943:Organophosphorus Compounds; D013438:Sulfhydryl Compounds; D013644:Tartrates; C015441:phospho-L-arginine; D019298:Glucose-6-Phosphate; D001120:Arginine; D013773:Tetramisole; D006867:Hydrolases; D000469:Alkaline Phosphatase; D010744:Phosphoric Monoester Hydrolases; D005952:Glucose-6-Phosphatase; C049021:phosphoamidase; C029768:tartaric acid