Surface lipoprotein sorting by crosstalk between Lpt and Lol pathways in gram-negative bacteria

Abstract Lipopolysaccharide (LPS) and lipoprotein, two essential components of the outer membrane (OM) in Gram-negative bacteria, play critical roles in bacterial physiology and pathogenicity. LPS translocation to the OM is mediated by LptDE, yet how lipoproteins sort to the cell surface remains elu...

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Main Authors: Qingshan Luo, Chengai Wang, Shuai Qiao, Shan Yu, Lianwan Chen, Seonghoon Kim, Kun Wang, Jiangge Zheng, Yong Zhang, Fan Wu, Xiaoguang Lei, Jizhong Lou, Michael Hennig, Wonpil Im, Long Miao, Min Zhou, Weiwei Bei, Yihua Huang
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-59660-y
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author Qingshan Luo
Chengai Wang
Shuai Qiao
Shan Yu
Lianwan Chen
Seonghoon Kim
Kun Wang
Jiangge Zheng
Yong Zhang
Fan Wu
Xiaoguang Lei
Jizhong Lou
Michael Hennig
Wonpil Im
Long Miao
Min Zhou
Weiwei Bei
Yihua Huang
author_facet Qingshan Luo
Chengai Wang
Shuai Qiao
Shan Yu
Lianwan Chen
Seonghoon Kim
Kun Wang
Jiangge Zheng
Yong Zhang
Fan Wu
Xiaoguang Lei
Jizhong Lou
Michael Hennig
Wonpil Im
Long Miao
Min Zhou
Weiwei Bei
Yihua Huang
author_sort Qingshan Luo
collection DOAJ
description Abstract Lipopolysaccharide (LPS) and lipoprotein, two essential components of the outer membrane (OM) in Gram-negative bacteria, play critical roles in bacterial physiology and pathogenicity. LPS translocation to the OM is mediated by LptDE, yet how lipoproteins sort to the cell surface remains elusive. Here, we identify candidate lipoproteins that may be transported to the cell surface via LptDE. Notably, we determine the crystal structures of LptDE from Pseudomonas aeruginosa and its complex with an endogenous Escherichia coli lipoprotein LptM. The paLptDE-LptM structure demonstrates that LptM may translocate to the OM via LptDE, in a manner similar to LPS transport. The β-barrel domain serves as a passage for the proteinaceous moiety while its acyl chains are transported outside. Our finding has been corroborated by results from native mass spectrometry, immunofluorescence, and photocrosslinking assays, revealing a potential surface exposed lipoproteins (SLPs) transport mechanism through which lipoproteins are loaded into LptA by LolCDE prior to assembly of the LptB2FGCADE complex. These observations provide initial evidence of functional overlap between the Lpt and Lol pathways, potentially broadening current perspectives on lipoprotein sorting.
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spelling doaj-art-d0dd36f8cf5b4874a318d30e2ae92c9b2025-08-20T01:53:19ZengNature PortfolioNature Communications2041-17232025-05-0116111410.1038/s41467-025-59660-ySurface lipoprotein sorting by crosstalk between Lpt and Lol pathways in gram-negative bacteriaQingshan Luo0Chengai Wang1Shuai Qiao2Shan Yu3Lianwan Chen4Seonghoon Kim5Kun Wang6Jiangge Zheng7Yong Zhang8Fan Wu9Xiaoguang Lei10Jizhong Lou11Michael Hennig12Wonpil Im13Long Miao14Min Zhou15Weiwei Bei16Yihua Huang17National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesNational Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesNational Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesNational Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesNational Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesDepartments of Biological Sciences and Bioengineering, Lehigh UniversityInstitute of Bio-analytical Chemistry, School of Chemical Engineering, Nanjing University of Science and TechnologyNational Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesKey Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesSynthetic and Functional Biomolecules Center, Beijing National Laboratory for Molecular Sciences, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, Peking UniversitySynthetic and Functional Biomolecules Center, Beijing National Laboratory for Molecular Sciences, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, Peking UniversityUniversity of Chinese Academy of SciencesleadXpro AG, Park InnovaareDepartments of Biological Sciences and Bioengineering, Lehigh UniversityUniversity of Chinese Academy of SciencesInstitute of Bio-analytical Chemistry, School of Chemical Engineering, Nanjing University of Science and TechnologyNational Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesNational Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of SciencesAbstract Lipopolysaccharide (LPS) and lipoprotein, two essential components of the outer membrane (OM) in Gram-negative bacteria, play critical roles in bacterial physiology and pathogenicity. LPS translocation to the OM is mediated by LptDE, yet how lipoproteins sort to the cell surface remains elusive. Here, we identify candidate lipoproteins that may be transported to the cell surface via LptDE. Notably, we determine the crystal structures of LptDE from Pseudomonas aeruginosa and its complex with an endogenous Escherichia coli lipoprotein LptM. The paLptDE-LptM structure demonstrates that LptM may translocate to the OM via LptDE, in a manner similar to LPS transport. The β-barrel domain serves as a passage for the proteinaceous moiety while its acyl chains are transported outside. Our finding has been corroborated by results from native mass spectrometry, immunofluorescence, and photocrosslinking assays, revealing a potential surface exposed lipoproteins (SLPs) transport mechanism through which lipoproteins are loaded into LptA by LolCDE prior to assembly of the LptB2FGCADE complex. These observations provide initial evidence of functional overlap between the Lpt and Lol pathways, potentially broadening current perspectives on lipoprotein sorting.https://doi.org/10.1038/s41467-025-59660-y
spellingShingle Qingshan Luo
Chengai Wang
Shuai Qiao
Shan Yu
Lianwan Chen
Seonghoon Kim
Kun Wang
Jiangge Zheng
Yong Zhang
Fan Wu
Xiaoguang Lei
Jizhong Lou
Michael Hennig
Wonpil Im
Long Miao
Min Zhou
Weiwei Bei
Yihua Huang
Surface lipoprotein sorting by crosstalk between Lpt and Lol pathways in gram-negative bacteria
Nature Communications
title Surface lipoprotein sorting by crosstalk between Lpt and Lol pathways in gram-negative bacteria
title_full Surface lipoprotein sorting by crosstalk between Lpt and Lol pathways in gram-negative bacteria
title_fullStr Surface lipoprotein sorting by crosstalk between Lpt and Lol pathways in gram-negative bacteria
title_full_unstemmed Surface lipoprotein sorting by crosstalk between Lpt and Lol pathways in gram-negative bacteria
title_short Surface lipoprotein sorting by crosstalk between Lpt and Lol pathways in gram-negative bacteria
title_sort surface lipoprotein sorting by crosstalk between lpt and lol pathways in gram negative bacteria
url https://doi.org/10.1038/s41467-025-59660-y
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