| Item type |
学術雑誌論文 / Journal Article_02(1) |
| 公開日 |
2008-02-05 |
| タイトル |
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タイトル |
Substrate specificity of Ca(2+)/calmodulin-dependent protein kinase phosphatase: kinetic studies using synthetic phosphopeptides as model substrates |
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言語 |
en |
| 言語 |
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言語 |
eng |
| 資源タイプ |
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資源タイプ |
journal article |
| 著者 |
石田, 敦彦
Shigeri, Y
Tatsu, Y
Endo, Y
Kameshita, I
Okuno, S
Kitani, T
Takeuchi, M
Yumoto, N
Fujisawa, H
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| 著者 ローマ字 |
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Ishida, Atsuhiko |
| 書誌情報 |
The Journal of Biochemistry
巻 129,
号 5,
p. 745-753,
発行日 2001-05-01
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| ISSN |
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収録物識別子タイプ |
ISSN |
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収録物識別子 |
0021-924X |
| リンクURL |
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内容記述タイプ |
Other |
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内容記述 |
http://jb.oxfordjournals.org/cgi/content/abstract/129/5/745 | http://jb.oxfordjournals.org/cgi/content/abstract/129/5/745 |
| 抄録 |
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内容記述タイプ |
Abstract |
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内容記述 |
Ca(2+)/calmodulin-dependent protein kinase phosphatase (CaMKPase) dephosphorylates and regulates multifunctional Ca(2+)/calmodulin-dependent protein kinases. In order to elucidate the mechanism of substrate recognition by CaMKPase, we chemically synthesized a variety of phosphopeptide analogs and carried out kinetic analysis using them as CaMKPase substrates. This is the first report using systematically synthesized phosphopeptides as substrates for kinetic studies on substrate specificities of protein Ser/Thr phosphatases. CaMKPase was shown to be a protein Ser/Thr phosphatase having a strong preference for a phospho-Thr residue. A Pro residue adjacent to the dephosphorylation site on the C-terminal side and acidic clusters around the dephosphorylation site had detrimental effects on dephosphorylation by CaMKPase. Deletion analysis of a model substrate peptide revealed that the minimal length of the substrate peptide was only 2 to 3 amino acid residues including the dephosphorylation site. The residues on the C-terminal side of the dephosphorylation site were not essential for dephosphorylation, whereas the residue adjacent to the dephosphorylation site on the N-terminal side was essential. Ala-scanning analysis suggested that CaMKPase did not recognize a specific motif around the dephosphorylation site. Myosin light chain phosphorylated by protein kinase C and Erk2 phosphorylated by MEK1 were poor substrates for CaMKPase, while a synthetic phosphopeptide corresponding to the sequence around the phosphorylation site of the former was not dephosphorylated by CaMKPase but that of the latter was fairly good substrate. These data suggest that substrate specificity of CaMKPase is determined by higher-order structure of the substrate protein rather than by the primary structure around its dephosphorylation site. Use of phosphopeptide substrates also revealed that poly-L-lysine, an activator for CaMKPase, activated the enzyme mainly through increase in the V(max) values. |
| 注記 |
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内容記述タイプ |
Other |
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注記 |
This is a pre-copy-editing, author-produced PDF of an article accepted for publication in Journal of Biochemistry following peer review. The definitive publisher-authenticated version Oxford University Press, Journal of Biochemistry, 129(5), 2001, 745-753 is available online at: http://jb.oxfordjournals.org/cgi/content/abstract/129/5/745.
\nauthor |
| 資源タイプ |
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内容記述タイプ |
Other |
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資源タイプ |
text |
| 著者版フラグ |
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出版タイプ |
AM |
| フォーマット |
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内容記述タイプ |
Other |
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内容記述 |
application/pdf |
| ID(XooNIps) |
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11328597 |
| 閲覧数(XooNIps) |
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| ダウンロード数(XooNIps) |
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2271 |
854.pdf (282.1 kB)
