Resolution, purification and characterization of the orthophosphate releasing activities from fracture callus calcifying cartilage.
Callus calcifying cartilage alkaline phosphatase was resolved by DEAE-cellulose column chromatography into two distinct phsophatase activities. The phosphatase activity which was eluted first from the column, (phosphatase I), was active towards a variety of phosphate esters, sodium pyrophosphatase and several linear polyphosphates, while the second phosphatase activity , (phosphatase II), was active toward simple phosphate esters but not towards sodium pyrophosphate and linear oligo or polyphosphates. All the phosphate esters, sodium pyrophosphate and polyphosphates at higher concentrations were inhibitory for phosphatase I. The modulating effects of magnesium, calcium, zinc and other phosphatase modulators have been investigated. Both phosphatases from callus calcifying cartilage were found to be substrates of neuraminidase with sialic acid as the product. Besides the difference in their specificity, the phosphatases were found to be immunologically different and to have different molecular weights, strong indication that they are different enzymes.
D000469:Alkaline Phosphatase / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D002146:Bony Callus / Q000201:enzymology*; D002113:Calcification, Physiologic; D002118:Calcium / Q000494:pharmacology; D002356:Cartilage / Q000201:enzymology*; D002499:Centrifugation, Density Gradient; D002848:Chromatography, DEAE-Cellulose; D004589:Electrophoresis, Disc; D050723:Fractures, Bone; D006863:Hydrogen-Ion Concentration; D005779:Immunodiffusion; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D009439:Neuraminidase; D011817:Rabbits; D012794:Sialic Acids / Q000032:analysis; D013329:Structure-Activity Relationship; D013977:Tibia / Q000201:enzymology; D013997:Time Factors; D014734:Vibrio cholerae / Q000201:enzymology; D015032:Zinc / Q000494:pharmacology
D012794:Sialic Acids; D000469:Alkaline Phosphatase; D009439:Neuraminidase; D008274:Magnesium; D015032:Zinc; D002118:Calcium