The phosphatidic acid pathway enzyme PlsX plays both catalytic and channeling roles in bacterial phospholipid synthesis
| العنوان: | The phosphatidic acid pathway enzyme PlsX plays both catalytic and channeling roles in bacterial phospholipid synthesis |
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| المؤلفون: | Frederico J. Gueiros-Filho, Caterina G. C. M. Netto, Diego E. Sastre, Diego de Mendoza, Marcos V.A.S. Navarro, Luis G.M. Basso, Jhonathan S. Benites Pariente, Daniela Albanesi, André A. Pulschen, Federico Machinandiarena |
| المصدر: | J Biol Chem Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | 0301 basic medicine, Substrate channeling, Phospholipid, Phosphatidic Acids, Biochemistry, Catalysis, 03 medical and health sciences, chemistry.chemical_compound, Bacterial Proteins, Escherichia coli, Lipid bilayer, Molecular Biology, Phospholipids, chemistry.chemical_classification, 030102 biochemistry & molecular biology, Lipogenesis, Fatty Acids, Fatty acid, Cell Biology, Phosphatidic acid, Lipids, 030104 developmental biology, Enzyme, chemistry, LIPÍDEOS, Acyltransferase, Membrane biogenesis, Metabolic Networks and Pathways, Bacillus subtilis |
| الوصف: | PlsX 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. |
| تدمد: | 1083-351X |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::6585af2c5badec3e7ae599a4bd8f59b4 https://pubmed.ncbi.nlm.nih.gov/31919098 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....6585af2c5badec3e7ae599a4bd8f59b4 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 1083351X |
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