Phosphate-binding tag : A new tool to visualize phosphorylated proteins

Molecular and Cellular Proteomics 5 巻 4 号 749-757 頁 2006-04 発行
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タイトル ( eng )
Phosphate-binding tag : A new tool to visualize phosphorylated proteins
作成者
Kinoshita-Kikuta Emiko
Takiyama Kei
収録物名
Molecular and Cellular Proteomics
5
4
開始ページ 749
終了ページ 757
抄録
We introduce two methods for the visualization of phosphorylated proteins using alkoxide-bridged dinuclear metal (i.e., Zn2+ or Mn2+) complexes as novel phosphate-binding tag (Phos-tag) molecules. Both Zn2+. and Mn2+.Phos-tag molecules preferentially capture phosphomonoester dianions bound to Ser, Thr, and Tyr residues. One method is based on an enhanced chemiluminescence (ECL) system using biotin-pendant Zn2+.Phos-tag and horseradish peroxidase-conjugated streptavidin (HRP.SA). We demonstrate the electroblotting analyses of protein phosphorylation status by the phosphate-selective ECL signals. Another method is based on the mobility shift of phosphorylated proteins in SDS-PAGE with polyacrylamide-bound Mn2+.Phos-tag. Phosphorylated proteins in the gel are visualized as slower migration bands compared with corresponding dephosphorylated proteins. We demonstrate the kinase and phosphatase assays by the phosphate-affinity electrophoresis (Mn2+.Phos-tag SDS-PAGE).
著者キーワード
Phosphoproteomics
Electroblotting analysis
Chemiluminescence detection
Phosphate-affinity electrophoresis
Phosphorylated protein
Phosphorylation
NDC分類
生物科学・一般生物学 [ 460 ]
言語
英語
資源タイプ 学術雑誌論文
出版者
American Society for Microbiology
発行日 2006-04
権利情報
Copyright (c) 2006 by the American Society for Biochemistry and Molecular Biology.
出版タイプ Author’s Original(十分な品質であるとして、著者から正式な査読に提出される版)
アクセス権 オープンアクセス
収録物識別子
[ISSN] 1535-9476
[DOI] 10.1074/mcp.T500024-MCP200
[NCID] AA11789524
[PMID] 16340016
[DOI] http://dx.doi.org/10.1074/mcp.T500024-MCP200 ~の異版である