磷(V)立体探针鉴定共价TLCD1抑制剂的蛋白质组配位性图

IF 15.6 1区 化学 Q1 CHEMISTRY, MULTIDISCIPLINARY
Hayden A. Sharma, Michael Bielecki, Meredith A. Holm, Ty M. Thompson, Yue Yin, Jacob B. Cravatt, Timothy B. Ware, Alex Reed, Molhm Nassir, Tamara El-Hayek Ewing, Bruno Melillo, J. Fernando Bazan, Phil S. Baran* and Benjamin F. Cravatt*, 
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引用次数: 0

摘要

基于活性的蛋白质分析(ABPP)立体异构定义的亲电化合物(“立体探针”)提供了一种通用的方法来发现天然生物系统中蛋白质的共价配体。本文报道了带有P(V)-草酸磷烷(OTP)反应基团的立体探针的合成和化学蛋白质组学表征。ABPP实验在人类癌细胞中发现了许多与OTP立体探针表现出立体选择反应性的蛋白质,并且我们证实了其中一些与重组蛋白的配体事件。OTP立体探针与特征不佳的跨膜蛋白TLCD1结合,破坏了细胞中单不饱和脂肪酸与磷脂酰乙醇胺脂质的结合,这种脂质组学表型反映了该蛋白的遗传破坏。利用AlphaFold2,我们发现TLCD1在结构上类似于辅酶a依赖性脂质加工酶的神经酰胺合成酶和脂肪酸延长酶家族。这种结构相似性包括催化组氨酸残基的保留,其突变阻断了重组TLCD1的OTP立体探针反应活性和脂质重塑活性。综上所述,这些数据表明TLCD1在细胞中起着脂质酰基转移酶的作用,而OTP立体探针可以抑制这种酶的活性。因此,我们的发现阐明了亲电化合物的化学蛋白质组学分析如何促进人类细胞中关键脂质代谢酶的功能注释和化学抑制。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Proteomic Ligandability Maps of Phosphorus(V) Stereoprobes Identify Covalent TLCD1 Inhibitors

Proteomic Ligandability Maps of Phosphorus(V) Stereoprobes Identify Covalent TLCD1 Inhibitors

Activity-based protein profiling (ABPP) of stereoisomerically defined sets of electrophilic compounds (‘stereoprobes’) offers a versatile way to discover covalent ligands for proteins in native biological systems. Here we report the synthesis and chemical proteomic characterization of stereoprobes bearing a P(V)-oxathiaphospholane (OTP) reactive group. ABPP experiments identified numerous proteins in human cancer cells that showed stereoselective reactivity with OTP stereoprobes, and we confirmed several of these liganding events with recombinant proteins. OTP stereoprobes engaging the poorly characterized transmembrane protein TLCD1 impaired the incorporation of monounsaturated fatty acids into phosphatidylethanolamine lipids in cells, a lipidomic phenotype that mirrored genetic disruption of this protein. Using AlphaFold2, we found that TLCD1 structurally resembles the ceramide synthase and fatty acid elongase families of coenzyme A-dependent lipid processing enzymes. This structural similarity included conservation of catalytic histidine residues, the mutation of which blocked the OTP stereoprobe reactivity and lipid remodeling activity of recombinant TLCD1. Taken together, these data indicate that TLCD1 acts as a lipid acyltransferase in cells, and that OTP stereoprobes function as inhibitors of this enzymatic activity. Our findings thus illuminate how the chemical proteomic analysis of electrophilic compounds can facilitate the functional annotation and chemical inhibition of a key lipid metabolic enzyme in human cells.

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来源期刊
CiteScore
24.40
自引率
6.00%
发文量
2398
审稿时长
1.6 months
期刊介绍: The flagship journal of the American Chemical Society, known as the Journal of the American Chemical Society (JACS), has been a prestigious publication since its establishment in 1879. It holds a preeminent position in the field of chemistry and related interdisciplinary sciences. JACS is committed to disseminating cutting-edge research papers, covering a wide range of topics, and encompasses approximately 19,000 pages of Articles, Communications, and Perspectives annually. With a weekly publication frequency, JACS plays a vital role in advancing the field of chemistry by providing essential research.
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