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基于机制的陷可以发现蛋白酶和酶基质

Shan Tang¹, Adam T Beattie², Lucie Kafkova³

¹Medical Research Council Laboratory of Molecular Biology, Cambridge, UK. stang@mrc-lmb.cam.ac.uk.

|February 17, 2022

概括

研究人员开发了新的光激活基质陷,以识别未知的酶,包括蛋白酶的功能. 这种方法揭示了人类细胞中的新蛋白质酶基质和酶活性.

科学领域:

生物化学
分子生物学
酵素学

背景情况:

包括蛋白酶在内的酶在生物过程中至关重要,其中很大一部分是药物点.
许多水溶酶,特别是膜结合蛋白酶的特异性活动和基质偏好在很大程度上未被描述.
了解这些酶对于药物发现和解读细胞机制至关重要.

研究的目的:

开发一种用于识别酶基质和活性的新策略.
在复杂的生物系统中创建基于机制的光激活基板陷.
发现非特征蛋白酶和氨酸水溶酶的新基质和功能.

主要方法:

基因编码的2,3-diaminopropionic酸替代了水溶酶中的催化核 (氨酸或氨酸).
开发的光激活基质陷,形成稳定的乙酶中间体以捕获基质.
在复杂的混合物和活哺乳动物细胞中应用这些陷以确定酶基质相互作用.

主要成果:

已确定蛋白质酶的新基质,包括膜内蛋白质酶RHBDL4 (含有Rhomboid Family Domain 4).
证明了RHBDL4在散射ER类型I跨膜蛋白的光片段和促进伴侣分泌中的作用.
描述了视网膜母细胞瘤结合蛋白9作为一种偏好芳香氨基酸分裂的氨基酶.

结论:

开发的基质陷策略为发现酶基质和活动提供了强大的范式.
这种方法可以在复杂的生物环境中对以前未知的酶进行功能性表征.
这些发现扩大了我们对蛋白质酶功能的理解,特别是关于膜蛋白和蛋白质分泌途径.

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