title: Quantitative Lipoproteomics in Clostridium difficile Reveals a Role for Lipoproteins in Sporulation.
abstract: Bacterial lipoproteins are surface exposed, anchored to the membrane by S-diacylglyceryl modification of the N-terminal cysteine thiol. They play important roles in many essential cellular processes and in bacterial pathogenesis. For example, Clostridium difficile is a Gram-positive anaerobe that causes severe gastrointestinal disease; however, its lipoproteome remains poorly characterized. Here we describe the application of metabolic tagging with alkyne-tagged lipid analogs, in combination with quantitative proteomics, to profile protein lipidation across diverse C. difficile strains and on inactivation of specific components of the lipoprotein biogenesis pathway. These studies provide the first comprehensive map of the C. difficile lipoproteome, demonstrate the existence of two active lipoprotein signal peptidases, and provide insights into lipoprotein function, implicating the lipoproteome in transmission of this pathogen.
authors.affiliation: Department of Chemistry, Imperial College London, London SW7 2AZ, UK; Life Sciences, Imperial College London, London SW7 2AZ, UK; Institute of Chemical Biology, Imperial College London, London SW7 2AZ, UK.
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Charlton
authors.affiliation: Department of Biosciences, College of Life and Environmental Sciences, University of Exeter, Exeter EX4 4QD, UK.
authors.forename: Andrea
authors.initials: A
authors.lastname: Kovacs-Simon
authors.affiliation: Department of Biosciences, College of Life and Environmental Sciences, University of Exeter, Exeter EX4 4QD, UK.
authors.forename: Stephen L
authors.initials: SL
authors.lastname: Michell
authors.affiliation: Life Sciences, Imperial College London, London SW7 2AZ, UK; Institute of Chemical Biology, Imperial College London, London SW7 2AZ, UK. Electronic address: n.fairweather@imperial.ac.uk.
authors.forename: Neil F
authors.initials: NF
authors.lastname: Fairweather
authors.affiliation: Department of Chemistry, Imperial College London, London SW7 2AZ, UK; Institute of Chemical Biology, Imperial College London, London SW7 2AZ, UK. Electronic address: e.tate@imperial.ac.uk.
authors.forename: Edward W
authors.initials: EW
authors.lastname: Tate
mesh_terms: D000480:Alkynes / Q000737:chemistry; D055162:Biocatalysis; D002851:Chromatography, High Pressure Liquid; D016360:Clostridioides difficile / Q000502:physiology*; D008074:Lipoproteins / Q000737:chemistry / Q000378:metabolism*; D019814:Myristic Acid / Q000737:chemistry; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D013171:Spores, Bacterial / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D000480:Alkynes; D008074:Lipoproteins; D020543:Proteome; D019814:Myristic Acid