气候变化领域集成服务门户(CCPortal): Supramolecular assembly of the Escherichia coli LdcI upon acid stress

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DOI	10.1073/pnas.2014383118
	Supramolecular assembly of the Escherichia coli LdcI upon acid stress
	Jessop M.; Liesche C.; Felix J.; Desfosses A.; Baulard M.; Adam V.; Fraudeau A.; Huard K.; Effantin G.; Kleman J.-P.; Bacia-Verloop M.; Bourgeois D.; Gutsche I.
发表日期	2021
ISSN	00278424
卷号	118 期号:2
英文摘要	Pathogenic and commensal bacteria often have to resist the harsh acidity of the host stomach. The inducible lysine decarboxylase LdcI buffers the cytosol and the local extracellular environment to ensure enterobacterial survival at low pH. Here, we investigate the acid stress-response regulation of Escherichia coli LdcI by combining biochemical and biophysical characterization with negative stain and cryoelectron microscopy (cryo-EM) and wide-field and superresolution fluorescence imaging. Due to deleterious effects of fluorescent protein fusions on native LdcI decamers, we opt for three-dimensional localization of nanobody-labeled endogenous wild-type LdcI in acid-stressed E. coli cells and show that it organizes into distinct patches at the cell periphery. Consistent with recent hypotheses that in vivo clustering of metabolic enzymes often reflects their polymerization as a means of stimulus-induced regulation, we show that LdcI assembles into filaments in vitro at physiologically relevant low pH. We solve the structures of these filaments and of the LdcI decamer formed at neutral pH by cryo-EM and reveal the molecular determinants of LdcI polymerization, confirmed by mutational analysis. Finally, we propose a model for LdcI function inside the enterobacterial cell, providing a structural and mechanistic basis for further investigation of the role of its supramolecular organization in the acid stress response. © This open access article is distributed under Creative Commons Attribution-NonCommercialNoDerivatives License 4.0 (CC BY-NC-ND).
英文关键词	Cryo-EM; LdcI; Lysine decarboxylase; SMLM
语种	英语
scopus关键词	acidity; Article; bacterial cell; bacterial gene; bacterial virulence; biochemical analysis; biophysics; controlled study; cryoelectron microscopy; enzyme metabolism; Escherichia coli; fluorescence analysis; gene overexpression; immunofluorescence; immunohistochemistry; in vivo study; LdcI gene; molecular dynamics; mutational analysis; nonhuman; pH; polymerization; priority journal; protein interaction; protein localization; protein modification; supramolecular chemistry; wild type
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/181053
作者单位	Institut de Biologie Structurale (IBS), Université Grenoble Alpes (UGA), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), CNRS, Grenoble, F-38044, France
推荐引用方式 GB/T 7714	Jessop M.,Liesche C.,Felix J.,et al. Supramolecular assembly of the Escherichia coli LdcI upon acid stress[J],2021,118(2).
APA	Jessop M..,Liesche C..,Felix J..,Desfosses A..,Baulard M..,...&Gutsche I..(2021).Supramolecular assembly of the Escherichia coli LdcI upon acid stress.Proceedings of the National Academy of Sciences of the United States of America,118(2).
MLA	Jessop M.,et al."Supramolecular assembly of the Escherichia coli LdcI upon acid stress".Proceedings of the National Academy of Sciences of the United States of America 118.2(2021).