サイト・スペシフィック・ラベルされたLin28を用いた結合アッセイを用いたタンパク質-マイクロRNA相互作用の小分子阻害剤の発見
Donghyun Lim1, Wan Gi Byun2, Ja Young Koo2
1Department of Biophysics and Chemical Biology, Seoul National University , Seoul 08826, Korea.
Journal of the American Chemical Society
|September 27, 2016
まとめ
研究者らは Lin28-let-7の相互作用を阻害する小さな分子を特定しました この阻害剤は腫瘍抑制の let-7 miRNAの生成をがん細胞で促進し,新たながん治療の可能性を秘めています.
科学分野:
- 生物化学
- 分子生物学
- 腫瘍学
背景:
- マイクロRNA (miRNA) は遺伝子発現を調節し,細胞プロセスにおいて極めて重要です.
- let-7 miRNAファミリーは腫瘍抑制剤として作用するが,がんではしばしばダウンレギュレーションされる.
- リン28タンパク質はlet-7 miRNAの成熟を阻害し,がんの発生に寄与する.
研究 の 目的:
- Lin28- let-7の相互作用を阻害するものを発見するための高通量スクリーニングシステムを開発する.
- 腫瘍性リン28- レット7結合を妨げる小さな分子を特定する.
主な方法:
- フォースター共鳴エネルギー転送 (FRET) ベースのスクリーニングシステムを使用した.
- Lin28のサイト固有の光ラベリングのための非自然なアミノ酸変異とバイオオートホーゴン化学を用いた.
- 抑制剤を検知するために直接結合測定を行った.
主要な成果:
- Lin28- let-7の相互作用のための堅牢で信頼性の高いFRETベースの測定法を開発しました.
- Lin28- let-7相互作用の小分子阻害剤を特定した.
- リン28を発現する癌細胞におけるlet-7 miRNAの産生を増加させることが示された.
- let-7標的腫瘍遺伝子の減少を示した.
結論:
- 開発されたFRETアッセイは,Lin28- let-7相互作用の阻害剤を特定するのに有効です.
- 特定された小分子は,Let-7 miRNAのレベルを回復し,がん細胞における腫瘍遺伝子の発現を抑制することで,治療的可能性を示しています.
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