biostudies-literatureChristensen PMAmerican Heart AssociationCecil and Ida Green FoundationNIAID NIH HHSNational Institutes of HealthNIH HHSNIGMS NIH HHSNational Science FoundationRP94929https://www.ebi.ac.uk/biostudies/studies/S-EPMC11630815biostudies-literatureUnknown13Bacterial membranes are complex and dynamic, arising from an array of evolutionary pressures. One enzyme that alters membrane compositions through covalent lipid modification is MprF. We recently identified that <i>Streptococcus agalactiae</i> MprF synthesizes lysyl-phosphatidylglycerol (Lys-PG) from anionic PG, and a novel cationic lipid, lysyl-glucosyl-diacylglycerol (Lys-Glc-DAG), from neutral glycolipid Glc-DAG. This unexpected result prompted us to investigate whether Lys-Glc-DAG occurs in other MprF-containing bacteria, and whether other novel MprF products exist. Here, we studied protein sequence features determining MprF substrate specificity. First, pairwise analyses identified several streptococcal MprFs synthesizing Lys-Glc-DAG. Second, a restricted Boltzmann machine-guided appreLifeLipid discovery enabled by sequence statistics and machine learning.PMC1163081510.58275/AHA.23POST1013835.pc.gr.161273R01 AI178692R35 GM133631R35GM133631R01AI178692MCB-1943442R01 AI148366Uppuluri AMorcos FGuan ZChristensen PMMartin JJoyce LRPalmer KLWei YfalseLipid discovery enabled by sequence statistics and machine learning.Bacterial membranes are complex and dynamic, arising from an array of evolutionary pressures. One enzyme that alters membrane compositions through covalent lipid modification is MprF. We recently identified that <i>Streptococcus agalactiae</i> MprF synthesizes lysyl-phosphatidylglycerol (Lys-PG) from anionic PG, and a novel cationic lipid, lysyl-glucosyl-diacylglycerol (Lys-Glc-DAG), from neutral glycolipid Glc-DAG. This unexpected result prompted us to investigate whether Lys-Glc-DAG occurs in other MprF-containing bacteria, and whether other novel MprF products exist. Here, we studied protein sequence features determining MprF substrate specificity. First, pairwise analyses identified several streptococcal MprFs synthesizing Lys-Glc-DAG. Second, a restricted Boltzmann machine-guided appr2024-01-01T00:00:00Z2024 Dec2026-07-15T08:03:16.531Z2025-04-19T14:36:49.657ZS-EPMC116308153965651610.7554/eLife.94929