Bioorthogonal Monomycolate of Trehalose Disclosed the O-Mycoloylation of Mycoloyltransferases and Other Cell Envelope Proteins in C. glutamicum.

IF 3.5 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
ACS Chemical Biology Pub Date : 2024-11-15 Epub Date: 2024-10-31 DOI:10.1021/acschembio.4c00502
Cécile Labarre, Yijie Zhang, Emilie Lesur, Marie Ley, Laila Sago, Christiane Dietrich, Célia de Sousa-d'Auria, Florence Constantinesco-Becker, Aurélie Baron, Gilles Doisneau, Dominique Urban, Guillaume Chevreux, Dominique Guianvarc'h, Yann Bourdreux, Nicolas Bayan
{"title":"Bioorthogonal Monomycolate of Trehalose Disclosed the <i>O</i>-Mycoloylation of Mycoloyltransferases and Other Cell Envelope Proteins in <i>C. glutamicum</i>.","authors":"Cécile Labarre, Yijie Zhang, Emilie Lesur, Marie Ley, Laila Sago, Christiane Dietrich, Célia de Sousa-d'Auria, Florence Constantinesco-Becker, Aurélie Baron, Gilles Doisneau, Dominique Urban, Guillaume Chevreux, Dominique Guianvarc'h, Yann Bourdreux, Nicolas Bayan","doi":"10.1021/acschembio.4c00502","DOIUrl":null,"url":null,"abstract":"<p><p>Protein mycoloylation is a recently identified unusual post-translational modification (PTM) exclusively observed in Mycobacteriales, an order of bacteria that includes several human pathogens. These bacteria possess a distinctive outer membrane, known as the mycomembrane, composed of very long-chain fatty acids called mycolic acids. It has been demonstrated that a few mycomembrane proteins undergo covalent modification with mycolic acids in the model organism <i>Corynebacterium glutamicum</i> through the action of mycoloyltransferase MytC. This PTM represents the first example of protein <i>O</i>-acylation in prokaryotes and also the first example of protein modification by mycolic acid. Many questions about the specificity of protein <i>O</i>-mycoloylation remain crucial for understanding its evolutionary significance in Mycobacteriales and its role in cell physiology. We have developed the first bioorthogonal mycolate donor featuring the natural mycolic acid pattern, enabling direct, unambiguous transfer of the lipid moiety to its acceptors and efficient metabolic labeling and enrichment of MytC protein substrates. Mass spectrometry analysis of the labeled proteins and comparative proteomic analysis of the cell envelope proteome between wild-type and Δ<i>mytC</i> strains identified an unbiased list of 21 proteins likely mycoloylated in the cell. The robustness of our approach is demonstrated by the successful biological validation of mycoloylation in 6 candidate proteins within wild-type cells, revealing the characteristic profile of proteins modified with natural mycolates. These findings provide interesting insights into the significance of this new lipidation pathway and pave the way for understanding their function, especially concerning the mycoloyltransferase family that includes the essential Antigen85 enzymes in Mycobacteria.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":"2359-2371"},"PeriodicalIF":3.5000,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Chemical Biology","FirstCategoryId":"88","ListUrlMain":"https://doi.org/10.1021/acschembio.4c00502","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/10/31 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

Protein mycoloylation is a recently identified unusual post-translational modification (PTM) exclusively observed in Mycobacteriales, an order of bacteria that includes several human pathogens. These bacteria possess a distinctive outer membrane, known as the mycomembrane, composed of very long-chain fatty acids called mycolic acids. It has been demonstrated that a few mycomembrane proteins undergo covalent modification with mycolic acids in the model organism Corynebacterium glutamicum through the action of mycoloyltransferase MytC. This PTM represents the first example of protein O-acylation in prokaryotes and also the first example of protein modification by mycolic acid. Many questions about the specificity of protein O-mycoloylation remain crucial for understanding its evolutionary significance in Mycobacteriales and its role in cell physiology. We have developed the first bioorthogonal mycolate donor featuring the natural mycolic acid pattern, enabling direct, unambiguous transfer of the lipid moiety to its acceptors and efficient metabolic labeling and enrichment of MytC protein substrates. Mass spectrometry analysis of the labeled proteins and comparative proteomic analysis of the cell envelope proteome between wild-type and ΔmytC strains identified an unbiased list of 21 proteins likely mycoloylated in the cell. The robustness of our approach is demonstrated by the successful biological validation of mycoloylation in 6 candidate proteins within wild-type cells, revealing the characteristic profile of proteins modified with natural mycolates. These findings provide interesting insights into the significance of this new lipidation pathway and pave the way for understanding their function, especially concerning the mycoloyltransferase family that includes the essential Antigen85 enzymes in Mycobacteria.

生物正交单酵素三卤糖揭示了谷氨酸棒状杆菌中霉菌酰转移酶和其他细胞包膜蛋白的 O-霉菌酰化。
蛋白质霉菌酰化是最近发现的一种不寻常的翻译后修饰(PTM),只在分枝杆菌(Mycobacteriales)中观察到。这些细菌有一种独特的外膜,称为霉菌膜,由称为霉酚酸的长链脂肪酸组成。研究表明,在模式生物谷氨酸棒杆菌(Corynebacterium glutamicum)中,通过霉菌酰基转移酶 MytC 的作用,一些霉菌膜蛋白质会与霉菌酸发生共价修饰。这种 PTM 是原核生物中蛋白质 O-酰化的第一个例子,也是霉菌醇酸修饰蛋白质的第一个例子。关于蛋白质 O-霉菌酰化的特异性的许多问题,对于理解其在分枝杆菌中的进化意义及其在细胞生理中的作用仍然至关重要。我们开发出了第一种具有天然霉菌酸模式的生物正交霉菌酸供体,可直接、明确地将脂质分子转移到其受体上,并对 MytC 蛋白底物进行高效的代谢标记和富集。对标记的蛋白质进行质谱分析,并对野生型菌株和ΔmytC 菌株的细胞包膜蛋白质组进行比较蛋白质组分析,确定了一份无偏见的 21 种可能在细胞中被霉菌酰化的蛋白质列表。野生型细胞中 6 个候选蛋白质的霉菌酰化成功地进行了生物学验证,揭示了天然霉菌酸盐修饰蛋白质的特征,从而证明了我们的方法的稳健性。这些发现为了解这一新的脂化途径的意义提供了有趣的见解,并为了解它们的功能铺平了道路,尤其是霉菌酰基转移酶家族的功能,该家族包括分枝杆菌中重要的抗原85酶。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
ACS Chemical Biology
ACS Chemical Biology 生物-生化与分子生物学
CiteScore
7.50
自引率
5.00%
发文量
353
审稿时长
3.3 months
期刊介绍: ACS Chemical Biology provides an international forum for the rapid communication of research that broadly embraces the interface between chemistry and biology. The journal also serves as a forum to facilitate the communication between biologists and chemists that will translate into new research opportunities and discoveries. Results will be published in which molecular reasoning has been used to probe questions through in vitro investigations, cell biological methods, or organismic studies. We welcome mechanistic studies on proteins, nucleic acids, sugars, lipids, and nonbiological polymers. The journal serves a large scientific community, exploring cellular function from both chemical and biological perspectives. It is understood that submitted work is based upon original results and has not been published previously.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信