title: Purification and characterization of alkaline phosphatase containing phosphotyrosyl phosphatase activity from the bacterium Prevotella intermedia.
abstract: A novel alkaline phosphatase, designated PiALP, has been purified and characterized from Prevotella intermedia ATCC 25611, an anaerobe implicated in progressive periodontal disease. The enzyme was a homodimer of apparently identical subunits of Mr 54 kDa. Thiol-reducing agents completely inhibited the purified enzyme. The enzyme was highly stable even at 80 degrees C. It exhibited substantial activity against tyrosine-phosphate-containing Raytide. The phosphatase activity was sensitive to orthovanadate and Zn2+ but highly resistant to okadaic acid. The amino acid sequence of peptides derived from PiALP showed a high degree of identity (65%) with alkaline phosphatases from Zymomonas mobilis and Synechococcus. The present results imply that PiALP might represent a new family of alkaline phosphotyrosyl phosphatases which has not been described previously.
authors.affiliation: Department of Preventive Dentistry, Kyushu Dental College, Kitakyushu, Japan. ansai@kyu-dent.ac.jp
authors.forename: T
authors.initials: T
authors.lastname: Ansai
authors.forename: S
authors.initials: S
authors.lastname: Awano
authors.forename: X
authors.initials: X
authors.lastname: Chen
authors.forename: T
authors.initials: T
authors.lastname: Fuchi
authors.forename: T
authors.initials: T
authors.lastname: Arimoto
authors.forename: S
authors.initials: S
authors.lastname: Akifusa
authors.forename: T
authors.initials: T
authors.lastname: Takehara
mesh_terms: D000469:Alkaline Phosphatase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006358:Hot Temperature; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D018713:Prevotella intermedia / Q000201:enzymology*; D017027:Protein Tyrosine Phosphatases / Q000302:isolation & purification / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D013816:Thermodynamics
chemical_list: D010446:Peptide Fragments; D000469:Alkaline Phosphatase; D017027:Protein Tyrosine Phosphatases