The mycobacterium lipid transporter MmpL3 is dimeric in detergent solution, SMALPs and reconstituted nanodiscs†

IF 4.2 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY RSC Chemical Biology Pub Date : 2024-07-29 DOI:10.1039/D4CB00110A
Sara Cioccolo, Joseph D. Barritt, Naomi Pollock, Zoe Hall, Julia Babuta, Pooja Sridhar, Alicia Just, Nina Morgner, Tim Dafforn, Ian Gould and Bernadette Byrne
{"title":"The mycobacterium lipid transporter MmpL3 is dimeric in detergent solution, SMALPs and reconstituted nanodiscs†","authors":"Sara Cioccolo, Joseph D. Barritt, Naomi Pollock, Zoe Hall, Julia Babuta, Pooja Sridhar, Alicia Just, Nina Morgner, Tim Dafforn, Ian Gould and Bernadette Byrne","doi":"10.1039/D4CB00110A","DOIUrl":null,"url":null,"abstract":"<p >The mycobacterial membrane protein large 3 (MmpL3) transports key precursor lipids to the outer membrane of Mycobacterium species. Multiple structures of MmpL3 from both <em>M. tuberculosis</em> and <em>M. smegmatis</em> in various conformational states indicate that the protein is both structurally and functionally monomeric. However, most other resistance, nodulation and cell division (RND) transporters structurally characterised to date are either dimeric or trimeric. Here we present an in depth biophysical and computational analysis revealing that MmpL3 from <em>M. smegmatis</em> exists as a dimer in a variety of membrane mimetic systems (SMALPs, detergent-based solution and nanodiscs). Sucrose gradient separation of MmpL3 populations from <em>M. smegmatis</em>, reconstituted into nanodiscs, identified monomeric and dimeric populations of the protein using laser induced liquid bead ion desorption (LILBID), a native mass spectrometry technique. Preliminary cryo-EM analysis confirmed that MmpL3 forms physiological dimers. Untargeted lipidomics experiments on membrane protein co-purified lipids revealed PE and PG lipid classes were predominant. Molecular dynamics (MD) simulations, in the presence of physiologically-relevant lipid compositions revealed the likely dimer interface.</p>","PeriodicalId":40691,"journal":{"name":"RSC Chemical Biology","volume":" 9","pages":" 901-913"},"PeriodicalIF":4.2000,"publicationDate":"2024-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2024/cb/d4cb00110a?page=search","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"RSC Chemical Biology","FirstCategoryId":"1085","ListUrlMain":"https://pubs.rsc.org/en/content/articlelanding/2024/cb/d4cb00110a","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

The mycobacterial membrane protein large 3 (MmpL3) transports key precursor lipids to the outer membrane of Mycobacterium species. Multiple structures of MmpL3 from both M. tuberculosis and M. smegmatis in various conformational states indicate that the protein is both structurally and functionally monomeric. However, most other resistance, nodulation and cell division (RND) transporters structurally characterised to date are either dimeric or trimeric. Here we present an in depth biophysical and computational analysis revealing that MmpL3 from M. smegmatis exists as a dimer in a variety of membrane mimetic systems (SMALPs, detergent-based solution and nanodiscs). Sucrose gradient separation of MmpL3 populations from M. smegmatis, reconstituted into nanodiscs, identified monomeric and dimeric populations of the protein using laser induced liquid bead ion desorption (LILBID), a native mass spectrometry technique. Preliminary cryo-EM analysis confirmed that MmpL3 forms physiological dimers. Untargeted lipidomics experiments on membrane protein co-purified lipids revealed PE and PG lipid classes were predominant. Molecular dynamics (MD) simulations, in the presence of physiologically-relevant lipid compositions revealed the likely dimer interface.

Abstract Image

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
分枝杆菌脂质转运体 MmpL3 在洗涤剂溶液、SMALPs 和重组纳米盘中是二聚体
分枝杆菌膜蛋白大 3(MmpL3)将关键的前体脂质转运到分枝杆菌的外膜。结核分枝杆菌和烟曲霉分枝杆菌的 MmpL3 在不同构象状态下的多个结构表明,该蛋白在结构上和功能上都是单体。然而,迄今为止,大多数其他抗性、结核和细胞分裂(RND)转运体的结构特征要么是二聚体,要么是三聚体。在此,我们进行了深入的生物物理和计算分析,发现在各种膜模拟系统(SMALPs、去垢剂溶液和纳米盘)中,来自烟草真菌的 MmpL3 是以二聚体形式存在的。利用激光诱导液珠离子解吸(LILBID)--一种原生质谱技术--对重组到纳米盘中的来自烟曲霉的 MmpL3 群体进行蔗糖梯度分离,确定了该蛋白质的单体和二聚体群体。初步的冷冻电镜分析证实,MmpL3 形成了生理二聚体。对膜蛋白共纯化脂质进行的非靶向脂质组学实验显示,PE 和 PG 类脂质占主导地位。分子动力学模拟显示,在存在生理相关脂质成分的情况下,可能存在二聚体界面。
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 去求助
来源期刊
CiteScore
6.10
自引率
0.00%
发文量
128
审稿时长
10 weeks
期刊最新文献
Cultivating the future leaders of chemical biology. Rational engineering of an antimalarial peptide with enhanced proteolytic stability and preserved parasite invasion inhibitory activity. A nanoengineered tandem nitroreductase: designing a robust prodrug-activating nanoreactor. A platform of ADAPTive scaffolds: development of CDR-H3 β-hairpin mimics into covalent inhibitors of the PD1/PDL1 immune checkpoint. Back cover
×
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
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 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 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1