title: The outer membrane of Gram-negative bacteria: lipid A isolation and characterization.
abstract: The isolation and characterization of the lipid A domain of lipopolysaccharide (LPS) are important methodologies utilized to gain understanding of the Gram-negative cell envelope. Here, we describe protocols often employed by our laboratory for small- and large-scale isolation of lipid A from bacterial cells. Additionally, we describe various methodologies including isolation of radiolabeled lipid A, thin layer chromatography, and various mass spectrometry methods. Tandem mass spectrometry is an integral tool for the structural characterization of lipid A molecules, and both coventional collision induced dissociation (CID) and new ultraviolet photodissociation (UVPD) methods are described.
authors.affiliation: Section of Molecular Genetics and Microbiology, The University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: Jessica V
authors.initials: JV
authors.lastname: Hankins
authors.affiliation: Department of Chemistry and Biochemistry, The University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: James A
authors.initials: JA
authors.lastname: Madsen
authors.affiliation: The Institute of Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: Brittany D
authors.initials: BD
authors.lastname: Needham
authors.affiliation: Department of Chemistry and Biochemistry, The University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: Jennifer S
authors.initials: JS
authors.lastname: Brodbelt
authors.affiliation: Section of Molecular Genetics and Microbiology, The University of Texas at Austin, Austin, TX 78712, USA.|The Institute of Cellular and Molecular Biology, The University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: M Stephen
authors.initials: MS
authors.lastname: Trent
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002240:Carbohydrate Sequence; D002855:Chromatography, Thin Layer; D008050:Lipid A / Q000737:chemistry / Q000302:isolation & purification*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data
chemical_list: D001425:Bacterial Outer Membrane Proteins; D008050:Lipid A