@article{oai:dmu.repo.nii.ac.jp:00000217,
 author = {清水, 左門 and 五十嵐, 吉彦 and Shimizu, Samon and Igarashi, Yoshihiko},
 issue = {1},
 journal = {Dokkyo journal of medical sciences},
 month = {Mar},
 note = {この実験の目的は破骨細胞から分泌される酒石酸抵抗性酸ホスファターゼ(tartrate-resistant acid phosphatase, TRACP)5bを特異的に定量する方法を発展させることである.ヘパリンカラムに吸着させた1mlの人血清はNaClの塩濃度勾配をかけることにより3つのTRACP活性に分かれた.最後のピークはATPase活性を持ち,TRACP5bと電気泳動位置が一致した.そのTRACP5bはおもに破骨細胞から分泌されたものと考えられている.このピークの高さは,骨形成期に最高値を示した後は,年齢の増加とともに減少した.なお,腎透析患者では血清TRACP5bは明らかに上昇した.二番目のピークはTRACP5aであった., Background and Aim : Mature osteoclasts are multinucleated giant cells that possess high levels of tartrate-resistant acid phosphatase (TRACP). The objective of our present study is to develop a column chromatographic method that separates isoforms of TRACP5 a and 5b in human serum. TRACP5b is considered to be derived from the osteoclast. Since no simple chromatographic method of measuring osteoclast-specific TRACP has been established, attempts were made to measure specifically TRACP isozymes in human serum using a heparin column. Methods : The serum was dialyzed against 20 mM Tris-HC1, 0.1 M NaCl, pH 7.2. After filtration, the sample was applied to a heparin column. The column was eluted with a linear gradient of sodium chloride from 0.1 to 1.05 M. TRACP activity was assayed with a colorimetric method. The final substrate concentration of p-nitrophenyl phosphate in the assay medium was 50 mM. The activity was measured in the presence of 40 mM sodium (+) tartrate with or without 45 mM sodium fluoride in lOOmM citrate buffer, pH 5.5. Results and Conclusion : Heparin column-bound TRACP in a human serum was clearly divided into three peaks when eluted with a linear gradient of sodium chloride. The last peak (peak III corresponded to TRACP5b which was first named according to its electrophoretic mobility by Lau et al and was considered to be secreted mainly from the osteoclast. The second peak (peak n) was found to be TRACP 5 a. Acid polyacrylamide gel electrophoresis showed that both peaks I and III corresponded to TRACP5b and the peak II to TRACP5a. Our method was not affected by hemolysis. In the sera of patients undergoing hemodialysis, the activity of TRACP5b was increased markedly. Our chromatographic method to separate TRACP5b can be useful to evaluate bone resorption., 原著, Original},
 pages = {45--55},
 title = {ヘパリンカラムによる血清由来の酒石酸抵抗性酸ホスファターゼの分析 : 骨吸収マーカー破骨細胞由来酒石酸抵抗性酸ホスファターゼをモニターする新しい方法},
 volume = {31},
 year = {2004},
 yomi = {シミズ, サモン and イガラシ, ヨシヒコ}
}