Tyrosine phosphatase-like activity of bone-type alkaline phosphatase at neutral pH range
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概要
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We aimed to determine physiological substrate and optimal pH for bone-type alkaline phosphatase(ALP, EC.3.1.3.1). Bone-type ALP was purified over 900-fold from clonal osteoblastic cells, MC3T3-E1, by butanol treatment, and chromatographies of anion exchange, Con A affinity and gel filtration. The purified enzyme showed a single protein band of molecular weight 76, 000 on SDS-polyacrylamide gel electrophoresis. The purified enzyme hydrolyzed p-nitrophenyl phosphate(pNPP)and dephosphorylated tyrosine-phosphorylated myelin basic protein(Tyr-P-MBP), but not serine-and threonine-phosphorylated MBP. Both activities were inhibited by levamisole, an ALP inhibitor, but not by okadaic acid or dephostatin(protein phosphatase inhibitors). The optimal pII ranges for pNPP and Tyr-P-MBP activities were 9.4 to 10.4 and 8.3 to 8.7, respectively. The ratios of activity at optimal pH to the one at pH 7 were 32.2 and 1.6 for pNPP and Tyr-P-MBP, respectively. Tyr-P-MBP dephosphorylation was competed by pNPP. These results suggest that bone-type ALP displays tyrosine phosphatase-like activity at neutral pH range and this activity may play an important role in physiological function of bone-type ALP.
- 硬組織再生生物学会の論文
著者
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MATSUMOTO Akira
Department of Microbiology, Kawasaki Medical School
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Sugawara Yuki
Department Of Oral Functional Science (section Of Orthodontics) Graduate School Of Dental Medicine H
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Sugawara Yuki
Department Of Dermatology Iwate Medical University School Of Medicine
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Koshikawa Mino
Department Of Oral Functional Science (section Of Orthodontics) Graduate School Of Dental Medicine H
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Koshikawa Mino
Department Of Orthodontics School Of Dentistry Hokkaido University
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Suzuki Kuniaki
Department Of Oral Pathobiological Science (section Of Dental Pharmacology) Graduate School Of Denta
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Iida Junichiro
Department Of Oral Functional Science (section Of Orthodontics) Graduate School Of Dental Medicine H
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Suzuki Kuniaki
Department Of Analytical Chemistry Tokyo College Of Pharmacy
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YOSHIKAWA Masatake
Department of Orthodontics School of Dentistry, Hokkaido University
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Yoshikawa Masatake
Department Of Oral Functional Science (section Of Orthodontics) Hokkaido University Graduate School
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Matsumoto Akira
Department Of Applied Chemistry
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Matsumoto Akira
Department Of Aeronautics And Astronautics Faculty Of Engineering Kyushu University
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