タンパク質の化学合成によって活性化されたデウビキキチナゼを標的とする
Shimrit Ohayon1, Liat Spasser, Amir Aharoni
1Department of Chemistry, Ben-Gurion University of the Negev, Beer Sheva 84105, Israel.
Journal of the American Chemical Society
|January 28, 2012
まとめ
研究者らは,デウビキチナゼ (DUB) 阻害剤をスクリーニングするために新しいFRETベースの測定法を開発しました. このアッセイでは,UCH-L3の強力な阻害剤を特定し,DUB関連疾患の薬剤発見を支援しました.
科学分野:
- バイオケミストリー バイオケミストリー
- 分子生物学は分子生物学である.
- ドラッグ・ディスカバリー・ドリッグ・ディスカバリー・ドリッグ・ディスカバリー・ドリッグ・ディスカバリー
背景:
- ユビキチネーションは,重要な細胞過程を調節する重要な翻訳後の修正である.
- デウビキチナゼ (Deubiquitinases,DUBs) はユビキチンを取り除き,細胞サイクル,発達,疾患に影響を及ぼします.
- DUBは治療目標ですが,薬剤発見は,アッセイの制限を含む課題に直面しています.
研究 の 目的:
- DUB阻害剤の高通量スクリーニングのための新しいFRETベースのアッセイの設計と検証.
- デウビキキチナゼの新型阻害剤を特定し,特にUCH-L3.3を標的とする.
主な方法:
- FRETペアで標識された化学合成ユビキチン化ペプチドを用いたFRETベースのアッセイを開発しました.
- 開発されたアッセイを使用して,UCH-L3酵素に対して1000の化合物のライブラリをスクリーニングしました.
- 特定された阻害剤の効能と作用機構を特徴づけた.
主要な成果:
- DUBスクリーニングのために,FRETベースの繊細な測定法を開発し,成功裏に実装しました.
- 特定の運動パラメータを持つUCH-L3の新規で強力な阻害剤を特定した (k (((inact) = 0.065 min ((-1),K ((i) = 0.8 μM).
- UCH-L1.1などの他のDUBに対するアッセイの適用性を実証した.
結論:
- 新しいFRETアッセイは,DUB阻害剤の効率的なスクリーニングを容易にする.
- 特定された強力なUCH-L3阻害剤は,治療開発のための有望な鉛化合物を表しています.
- この測定プラットフォームは,健康と病気におけるユビキチンシステムの役割に関する研究を支援しています.
関連する概念動画
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In this pathway, the target proteins are first tagged with small proteins called ubiquitin. This involves participation of a series of enzymes including— E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3 (ubiquitin...
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The Proteasome
Eukaryotic cells can degrade proteins through several pathways. One of the most important amongst these is the ubiquitin-proteasome pathway. It helps the cell eliminate the misfolded, damaged, or unwarranted cytoplasmic proteins in a highly specific manner.
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. A series of enzymes carry out the ubiquitination of the target proteins - E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...
In this pathway, the target proteins are first tagged with small proteins called ubiquitin. A series of enzymes carry out the ubiquitination of the target proteins - E1 (ubiquitin-activating enzyme), E2 (ubiquitin-conjugating enzyme), and E3...
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Protein degradation plays two important roles in the cells. It helps to protect cells from misfolded or damaged proteins before they lead to a...
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Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein.
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The Proteasome Structure
The ubiquitin-proteasome pathway is a well-known mechanism utilized by eukaryotic cells to remove cytoplasmic proteins that are misfolded, damaged, or no longer needed. In this pathway, the protein that needs to be eliminated undergoes a process called ubiquitination, where a chain of ubiquitin molecules is attached to the 48th lysine residue of the target protein. This ubiquitin modification helps the proteasome distinguish between a target protein and a healthy protein.
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