Purification and characterization of an acid phosphatase from Mycoplasma fermentans.
An acid phosphatase associated with the cell membranes of Mycoplasma fermentans was released from the membranes with Triton X-100, then purified by ion-exchange chromatography on DEAE-Sephacel and CM-Sepharose, followed by affinity chromatography on Con A-Sepharose. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme revealed a single band with a molecular mass of 31.2 kilodaltons. The enzyme activity toward p-nitrophenyl phosphate was enhanced remarkably by Cu2+, Co2+ and Mg2+, but the activity was not inhibited by EDTA. The enzyme dephosphorylated O-phospho-L-tyrosine as well as p-nitrophenyl phosphate, but not O-phospho-L-threonine, O-phospho-L-serine, glucose-1-phosphate, phosphoryl choline and adenosine triphosphate. The level of the O-phospho-L-tyrosine phosphatase activity was the highest in Mycoplasma faucium and the second highest in Mycoplasma fermentans of all tested human mycoplasmas.
Noda
M
M
Department of Oral Bacteriology, Hokkaido University School of Dentistry, Sapporo, Japan.
Shibata
K
K
Sawa
Y
Y
Shimokoube
H
H
Watanabe
T
T
D000135:Acid Phosphatase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004795:Enzyme Stability; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D016858:Mycoplasma fermentans / Q000201:enzymology* / Q000302:isolation & purification; D009596:Nitrophenols; D009943:Organophosphorus Compounds; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism; D013379:Substrate Specificity
D009596:Nitrophenols; D009943:Organophosphorus Compounds; C008644:nitrophenylphosphate; D000135:Acid Phosphatase; D017027:Protein Tyrosine Phosphatases