Chromatographic separation of two acid phosphatases from rat bone.
Extracts of tibiae of suckling rats were prepared with 0.3 M KCl containing 0.1% Triton X-100 and were chromatographed with CM-52 cellulose. Most of the acid phosphatase activity determined with p-nitrophenylphosphate (p-NPP) was bound to the cellulose and could be eluted with a sodium acetate buffer gradient in 2 distinct peaks. The major peak, E2, was bound strongly to the cellulose and showed high activity with p-NPP and inorganic pyrophosphate (P-Pi), but only slight activity with beta-glycerophosphate (beta-GP) and was unaffected by tartrate. The minor peak, E1, was weakly bound to the adsorbent, showed equal activity with p-NPP and beta-GP, but negligible activity with P-Pi and was completely inhibited by tartrate. These results support earlier evidence suggesting that bone contains at least 2 different acid phosphatases and that the more abundant enzyme may function as a pyrophosphatase.
D009597:4-Nitrophenylphosphatase / Q000378:metabolism; D000135:Acid Phosphatase / Q000302:isolation & purification* / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001842:Bone and Bones / Q000201:enzymology*; D002845:Chromatography; D011756:Diphosphates / Q000378:metabolism; D005994:Glycerophosphates / Q000378:metabolism; D009596:Nitrophenols; D009943:Organophosphorus Compounds / Q000378:metabolism; D011755:Pyrophosphatases / Q000378:metabolism; D051381:Rats; D013379:Substrate Specificity; D013644:Tartrates / Q000494:pharmacology; D013977:Tibia
D011756:Diphosphates; D005994:Glycerophosphates; D009596:Nitrophenols; D009943:Organophosphorus Compounds; D013644:Tartrates; D000469:Alkaline Phosphatase; D000135:Acid Phosphatase; D009597:4-Nitrophenylphosphatase; D011755:Pyrophosphatases