化合物ビーズによる化合物結合蛋白質の同定
概要
アガロースビーズ又は磁性ビーズに化合物アレイと同様に光親和型固定化法により化合物を固定化したビーズを作製する。化合物ビーズを用いて、細胞抽出液から化合物に結合する蛋白質を探索することができる。
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Identification of M-GFN-binding proteins.
(A) Model structure of M-GFN beads. (B) Detection of the coprecipitated proteins for M-GFN beads from RAW264 cell lysates. RAW264 cell lysates were precleared with control beads and incubated with M-GFN beads. The reacted beads were washed, and the eluted proteins were subjected to SDS/PAGE and visualized by CBB staining. The coprecipitated proteins for M-GFN beads were identified as described in Materials and Methods. (C-E) Purified His-tagged SGTA (C), GLO1 (D), or SCP2 (E) protein was incubated with control beads and M-GFN beads in the presence or absence of M-GFN as a competitor. The reacted beads were washed, and the eluted proteins were immunoblotted with anti-Xpress Ab, which allows detection of recombinant His-tagged proteins containing the N-terminal leader peptide (Xpress epitope). C, Control beads; M, M-GFN beads.(Ref:2)
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References:
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N. Kanoh, K. Honda, S. Simizu, M. Muroi, H. Osada.
Photo-crosslinked small molecule affinity matrix for facilitating forward and reverse chemical genetics.
Angew. Chem. Int. Ed., 44, 3559-3562 (2005).
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M. Kawatani, H. Okumura, K. Honda, N. Kanoh, M. Muroi, N. Dohmae, M. Takami, M. Kitagawa, Y. Futamura, M. Imoto, H. Osada.
The Identification of an osteoclastogenesis inhibitor through the inhibition of glyoxalase I.
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