title: The lipid raft proteome of Borrelia burgdorferi.
abstract: Eukaryotic lipid rafts are membrane microdomains that have significant amounts of cholesterol and a selective set of proteins that have been associated with multiple biological functions. The Lyme disease agent, Borrelia burgdorferi, is one of an increasing number of bacterial pathogens that incorporates cholesterol onto its membrane, and form cholesterol glycolipid domains that possess all the hallmarks of eukaryotic lipid rafts. In this study, we isolated lipid rafts from cultured B. burgdorferi as a detergent resistant membrane (DRM) fraction on density gradients, and characterized those molecules that partitioned exclusively or are highly enriched in these domains. Cholesterol glycolipids, the previously known raft-associated lipoproteins OspA and OpsB, and cholera toxin partitioned into the lipid rafts fraction indicating compatibility with components of the DRM. The proteome of lipid rafts was analyzed by a combination of LC-MS/MS or MudPIT. Identified proteins were analyzed in silico for parameters that included localization, isoelectric point, molecular mass and biological function. The proteome provided a consistent pattern of lipoproteins, proteases and their substrates, sensing molecules and prokaryotic homologs of eukaryotic lipid rafts. This study provides the first analysis of a prokaryotic lipid raft and has relevance for the biology of Borrelia, other pathogenic bacteria, as well as for the evolution of these structures. All MS data have been deposited in the ProteomeXchange with identifier PXD002365 (http://proteomecentral.proteomexchange.org/dataset/PXD002365).
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY, USA.
authors.forename: Alvaro
authors.initials: A
authors.lastname: Toledo
authors.affiliation: Laufer Center for Physical and Quantitative Biology, Stony Brook University, Stony Brook, NY, USA.
authors.forename: Alberto
authors.initials: A
authors.lastname: Pérez
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY, USA.|New York State Department of Health, Stony Brook University, Stony Brook, NY, USA.
authors.forename: James L
authors.initials: JL
authors.lastname: Coleman
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY, USA.
authors.forename: Jorge L
authors.initials: JL
authors.lastname: Benach
mesh_terms: D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000032:analysis*; D000954:Antigens, Surface / Q000032:analysis*; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis*; D001428:Bacterial Vaccines / Q000032:analysis*; D025065:Borrelia burgdorferi / Q000737:chemistry*; D002772:Cholera Toxin / Q000032:analysis*; D002853:Chromatography, Liquid; D003902:Detergents / Q000737:chemistry; D008074:Lipoproteins / Q000032:analysis*; D008193:Lyme Disease / Q000382:microbiology; D021962:Membrane Microdomains / Q000737:chemistry*; D008969:Molecular Sequence Data; D020543:Proteome / Q000032:analysis*; D016415:Sequence Alignment; D053719:Tandem Mass Spectrometry
chemical_list: D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D003902:Detergents; D008074:Lipoproteins; C072045:OspA protein; D020543:Proteome; C077030:OspB protein, Borrelia burgdorferi; D002772:Cholera Toxin
keywords: Borrelia; Lipid rafts; Lyme disease; Microbiology; Proteome