在p21激活酶5的中介激活状态中,通过阿菲默介导的锁定会导致酶抑制
Heather L Martin1, Amy L Turner2, Julie Higgins3
1BioScreening Technology Group, Leeds Institutes of Molecular Medicine, University of Leeds, Leeds LS9 7TF, UK; Division of Molecular Medicine, Leeds Institute of Medical Research at St James's University Hospital, University of Leeds, Leeds LS9 7TF, UK; School of Molecular and Cellular Biology, University of Leeds, Leeds LS2 9JT, UK.
Cell reports
|September 30, 2023
概括
研究人员发现了一种新的方法来抑制p21激活激酶5 (PAK5) 使用阿菲默试剂. 这一发现为针对尿路细胞癌的新酶抑制剂疗法开辟了可能性.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 癌症研究 癌症研究
背景情况:
- 激酶是关键的治疗点,抑制剂通过各种机制起作用,如ATP结合阻断或共价修饰.
- 21激活激酶5 (PAK5) 在尿路癌细胞存活和增殖中起作用.
- 了解激酶抑制机制是开发新疗法的关键.
研究的目的:
- 为了识别和描述一种新的激酶抑制机制.
- 探索PAK5在尿路癌中的作用.
- 开发 Affimer 试剂,用于研究酶功能和抑制.
主要方法:
- 在尿道癌细胞中进行了高含量的成像RNA干扰 (RNAi) 屏幕,以识别关键蛋白质.
- 开发并使用 Affimer 试剂来捕获中间蛋白质状态并识别抑制机制.
- 进行了联合结晶研究,以阐明 Affimer 和 PAK5.5 之间的结合相互作用.
主要成果:
- 抑制PAK5导致细胞数量减少,G1/S阶段停止,以及尿路细胞癌细胞的细胞扩大.
- 阿菲默试剂PAK5-Af17有效地重复了PAK5沉默观察到的表型.
- 同结晶显示,PAK5-Af17与PAK5的P+1口袋结合,稳定了部分激活的状态.
结论:
- PAK5对于尿路癌细胞系的存活和扩散至关重要.
- 可以使用affimer试剂来捕获中间状态中的激酶,揭示新的抑制机制.
- 结合P+1口袋代表了一类新的激酶抑制,与ATP竞争性或共价性抑制剂不同.
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