The phosphatidic acid pathway enzyme PlsX plays both catalytic and channeling roles in bacterial phospholipid synthesis

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dc.citation.titleJournal of Biological Chemistry (JBC)
dc.citation.volume295
dc.creatorSastre, Diego Emiliano
dc.creatorPulschen, André A.
dc.creatorBasso , Luis G.M.
dc.creatorBenites Pariente, Jhonathan S.
dc.creatorMarques Netto, Caterina G.C.
dc.creatorMachinandiarena, Federico
dc.creatorAlbanesi, Daniela
dc.creatorNavarro, Marcos V.A.S.
dc.creatorDe Mendoza, Diego
dc.creatorGueiros-Filho, Frederico J.
dc.date.accessioned2024-05-16T12:41:08Z
dc.date.available2024-05-16T12:41:08Z
dc.date.issued2020-01-09
dc.description.abstractPlsX is the first enzyme in the pathway that produces phosphatidic acid in Gram-positive bacteria. It makes acylphosphate from acyl-acyl carrier protein (acyl-ACP) and is also involved in coordinating phospholipid and fatty acid biosyntheses. PlsX is a peripheral membrane enzyme in Bacillus subtilis, but how it associates with the membrane remains largely unknown. In the present study, using fluorescence microscopy, liposome sedimentation, differential scanning calorimetry, and acyltransferase assays, we determined that PlsX binds directly to lipid bilayers and identified its membrane anchoring moiety, consisting of a hydrophobic loop located at the tip of two amphipathic dimerization helices. To establish the role of the membrane association of PlsX in acylphosphate synthesis and in the flux through the phosphatidic acid pathway, we then created mutations and gene fusions that prevent PlsX's interaction with the membrane. Interestingly, phospholipid synthesis was severely hampered in cells in which PlsX was detached from the membrane, and results from metabolic labeling indicated that these cells accumulated free fatty acids. Because the same mutations did not affect PlsX transacylase activity, we conclude that membrane association is required for the proper delivery of PlsX's product to PlsY, the next enzyme in the phosphatidic acid pathway. We conclude that PlsX plays a dual role in phospholipid synthesis, acting both as a catalyst and as a chaperone protein that mediates substrate channeling into the pathway.
dc.description.filFil: Sastre, Diego Emiliano. Universidade de São Paulo. Instituto de Química. Departamento de Bioquímica; Brasil.
dc.description.filFil: Sastre, Diego Emiliano. Universidade de São Paulo. Instituto de Física de São Carlos. Departamento de Biofísica Molecular. Grupo de Biofísica Molecular “Sergio Mascarenhas”; Brasil.
dc.description.filFil: Pulschen, André A. Universidade de São Paulo. Instituto de Química. Departamento de Bioquímica; Brasil.
dc.description.filFil: Basso, Luis G.M. Universidade de São Paulo. Faculdade de Filosofia Ciências e Letras de Ribeirão Preto. Departamento de Física; Brasil.
dc.description.filFil: Benites Pariente, Jhonathan S. Universidade de São Paulo. Instituto de Química. Departamento de Bioquímica; Brasil.
dc.description.filFil: Marques Netto, Caterina G.C. Universidade Federal de São Carlos. Departamento de Química; Brasil.
dc.description.filFil: Machinandiarena, Federico. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario (IBR-CONICET). Departamento de Microbiología; Argentina.
dc.description.filFil: Albanesi, Daniela. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario (IBR-CONICET). Departamento de Microbiología; Argentina.
dc.description.filFil: Navarro, Marcos V.A.S. Universidade de São Paulo. Instituto de Física de São Carlos. Departamento de Biofísica Molecular. Grupo de Biofísica Molecular “Sergio Mascarenhas”; Brasil.
dc.description.filFil: De Mendoza, Diego. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario (IBR-CONICET). Departamento de Microbiología; Argentina.
dc.description.filFil: Gueiros-Filho, Frederico J. Universidade de São Paulo. Instituto de Química. Departamento de Bioquímica; Brasil.
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP): grant 2016/05203-5, grant 2014/13411-1, grant 2015/21583-0, grant 2014/ 00206-0
dc.description.sponsorshipAgencia Nacional de Promoción de la Investigación, el Desarrollo Tecnológico y la Innovación (Agencia I+D+i): PICT 2016-1594
dc.description.versionpeerreviewed
dc.format.extent1-12
dc.identifier.e-issn1083-351X
dc.identifier.issn0021-9258
dc.identifier.urihttps://hdl.handle.net/2133/27045
dc.language.isoen
dc.publisherAmerican Society for Biochemistry and Molecular Biology
dc.relation.publisherversionhttps://doi.org/10.1074/jbc.ra119.011147
dc.relation.publisherversionhttps://www.sciencedirect.com/science/article/pii/S0021925817482895?via%3Dihub
dc.rightsopenAccess
dc.rights.holderSastre, Diego Emiliano
dc.rights.holderPulschen, André A.
dc.rights.holderBasso, Luis G.M.
dc.rights.holderBenites Pariente, Jhonathan S.
dc.rights.holderMarques Netto, Caterina G.C.
dc.rights.holderMachinandiarena, Federico
dc.rights.holderAlbanesi, Daniela
dc.rights.holderNavarro, Marcos V.A.S.
dc.rights.holderDe Mendoza, Diego
dc.rights.holderGueiros-Filho, Frederico J.
dc.rights.holderUniversidad Nacional de Rosario
dc.rights.textAttribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectGlycerophospholipid
dc.subjectGram-positive bacteria
dc.subjectAcyltransferase
dc.subjectMembrane biogenesis
dc.subjectLipid binding protein
dc.subjectAcylphosphate
dc.subjectPlsX
dc.subjectSubstrate channeling
dc.titleThe phosphatidic acid pathway enzyme PlsX plays both catalytic and channeling roles in bacterial phospholipid synthesis
dc.typearticulo
dc.type.versionpublishedVersion

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