Molecular Basis of Membrane Association by the Phosphatidylinositol Mannosyltransferase PimA Enzyme from Mycobacteria

Phosphatidyl-myo-inositol mannosyltransferase A (PimA) is an essential glycosyltransferase that initiates the biosynthetic pathway of phosphatidyl-myo-inositol mannoside, lipomannan, and lipoarabinomannan, which are key glycolipids/lipoglycans of the mycobacterial cell envelope. PimA belongs to a la...

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Detalles Bibliográficos
Autores: Rodrigo-Unzueta, Ane, Martínez, Mariano A., Comino, Natalia, Alzari, Pedro M., Chenal, Alexandre, Guerin, Marcelo E.
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2016
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/418340
Acceso en línea:http://hdl.handle.net/10261/418340
https://api.elsevier.com/content/abstract/scopus_id/84976898448
Access Level:acceso abierto
Palabra clave:Structure-function
Enzyme
Glycolipid
Glycosyltransferase
Membrane enzyme
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spelling Molecular Basis of Membrane Association by the Phosphatidylinositol Mannosyltransferase PimA Enzyme from MycobacteriaRodrigo-Unzueta, AneMartínez, Mariano A.Comino, NataliaAlzari, Pedro M.Chenal, AlexandreGuerin, Marcelo E.Structure-functionEnzymeGlycolipidGlycosyltransferaseMembrane enzymePhosphatidyl-myo-inositol mannosyltransferase A (PimA) is an essential glycosyltransferase that initiates the biosynthetic pathway of phosphatidyl-myo-inositol mannoside, lipomannan, and lipoarabinomannan, which are key glycolipids/lipoglycans of the mycobacterial cell envelope. PimA belongs to a large family of membrane-associated glycosyltransferases for which the understanding of the molecular mechanism and conformational changes that govern substrate/membrane recognition and catalysis remains a major challenge. Here, we determined that PimA preferentially binds to negatively charged phosphatidyl-myo-inositol substrate and non-substrate membrane model systems (small unilamellar vesicle) through its N-terminal domain, inducing an important structural reorganization of anionic phospholipids. By using a combination of single-point mutagenesis, circular dichroism, and a variety of fluorescence spectroscopy techniques, we determined that this interaction is mainly mediated by an amphipathic α-helix (α2), which undergoes a substantial conformational change and localizes in the vicinity of the negatively charged lipid headgroups and the very first carbon atoms of the acyl chains, at the PimA-phospholipid interface. Interestingly, a flexible region within the N-terminal domain, which undergoes β-strand-to-α-helix and α-helix-to-β-strand transitions during catalysis, interacts with anionic phospholipids; however, the effect is markedly less pronounced to that observed for the amphipathic α2, likely reflecting structural plasticity/variability. Altogether, we propose a model in which conformational transitions observed in PimA might reflect a molten globule state that confers to PimA, a higher affinity toward the dynamic and highly fluctuating lipid bilayer.This work was supported by the European Commission Contract HEALTH-F3-2011-260872, the Spanish Ministry of Economy and Competitiveness Contract BIO2013-49022-C2-2-R, the Basque Government (to M. E. G.), and Institut Pasteur (to A. C. and P. M. A.).Peer reviewedElsevierEuropean CommissionMinisterio de Economía y Competitividad (España)Eusko JaurlaritzaInstitut PasteurGuerin, Marcelo E. [0000-0001-9524-3184]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202620262016info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/418340https://api.elsevier.com/content/abstract/scopus_id/84976898448reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/FP7/260872/info:eu-repo/grantAgreement/MINECO//BIO2013-49022-C2-2-Rhttps://doi.org/10.1074/jbc.M116.723676Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/4183402026-05-22T06:33:51Z
dc.title.none.fl_str_mv Molecular Basis of Membrane Association by the Phosphatidylinositol Mannosyltransferase PimA Enzyme from Mycobacteria
title Molecular Basis of Membrane Association by the Phosphatidylinositol Mannosyltransferase PimA Enzyme from Mycobacteria
spellingShingle Molecular Basis of Membrane Association by the Phosphatidylinositol Mannosyltransferase PimA Enzyme from Mycobacteria
Rodrigo-Unzueta, Ane
Structure-function
Enzyme
Glycolipid
Glycosyltransferase
Membrane enzyme
title_short Molecular Basis of Membrane Association by the Phosphatidylinositol Mannosyltransferase PimA Enzyme from Mycobacteria
title_full Molecular Basis of Membrane Association by the Phosphatidylinositol Mannosyltransferase PimA Enzyme from Mycobacteria
title_fullStr Molecular Basis of Membrane Association by the Phosphatidylinositol Mannosyltransferase PimA Enzyme from Mycobacteria
title_full_unstemmed Molecular Basis of Membrane Association by the Phosphatidylinositol Mannosyltransferase PimA Enzyme from Mycobacteria
title_sort Molecular Basis of Membrane Association by the Phosphatidylinositol Mannosyltransferase PimA Enzyme from Mycobacteria
dc.creator.none.fl_str_mv Rodrigo-Unzueta, Ane
Martínez, Mariano A.
Comino, Natalia
Alzari, Pedro M.
Chenal, Alexandre
Guerin, Marcelo E.
author Rodrigo-Unzueta, Ane
author_facet Rodrigo-Unzueta, Ane
Martínez, Mariano A.
Comino, Natalia
Alzari, Pedro M.
Chenal, Alexandre
Guerin, Marcelo E.
author_role author
author2 Martínez, Mariano A.
Comino, Natalia
Alzari, Pedro M.
Chenal, Alexandre
Guerin, Marcelo E.
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv European Commission
Ministerio de Economía y Competitividad (España)
Eusko Jaurlaritza
Institut Pasteur
Guerin, Marcelo E. [0000-0001-9524-3184]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Structure-function
Enzyme
Glycolipid
Glycosyltransferase
Membrane enzyme
topic Structure-function
Enzyme
Glycolipid
Glycosyltransferase
Membrane enzyme
description Phosphatidyl-myo-inositol mannosyltransferase A (PimA) is an essential glycosyltransferase that initiates the biosynthetic pathway of phosphatidyl-myo-inositol mannoside, lipomannan, and lipoarabinomannan, which are key glycolipids/lipoglycans of the mycobacterial cell envelope. PimA belongs to a large family of membrane-associated glycosyltransferases for which the understanding of the molecular mechanism and conformational changes that govern substrate/membrane recognition and catalysis remains a major challenge. Here, we determined that PimA preferentially binds to negatively charged phosphatidyl-myo-inositol substrate and non-substrate membrane model systems (small unilamellar vesicle) through its N-terminal domain, inducing an important structural reorganization of anionic phospholipids. By using a combination of single-point mutagenesis, circular dichroism, and a variety of fluorescence spectroscopy techniques, we determined that this interaction is mainly mediated by an amphipathic α-helix (α2), which undergoes a substantial conformational change and localizes in the vicinity of the negatively charged lipid headgroups and the very first carbon atoms of the acyl chains, at the PimA-phospholipid interface. Interestingly, a flexible region within the N-terminal domain, which undergoes β-strand-to-α-helix and α-helix-to-β-strand transitions during catalysis, interacts with anionic phospholipids; however, the effect is markedly less pronounced to that observed for the amphipathic α2, likely reflecting structural plasticity/variability. Altogether, we propose a model in which conformational transitions observed in PimA might reflect a molten globule state that confers to PimA, a higher affinity toward the dynamic and highly fluctuating lipid bilayer.
publishDate 2016
dc.date.none.fl_str_mv 2016
2026
2026
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/418340
https://api.elsevier.com/content/abstract/scopus_id/84976898448
url http://hdl.handle.net/10261/418340
https://api.elsevier.com/content/abstract/scopus_id/84976898448
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/FP7/260872/
info:eu-repo/grantAgreement/MINECO//BIO2013-49022-C2-2-R
https://doi.org/10.1074/jbc.M116.723676
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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