title: Expression, refolding, and initial structural characterization of the Y. pestis Ail outer membrane protein in lipids.
abstract: Ail is an outer membrane protein and virulence factor of Yersinia pestis, an extremely pathogenic, category A biothreat agent, responsible for precipitating massive human plague pandemics throughout history. Due to its key role in bacterial adhesion to host cells and bacterial resistance to host defense, Ail is a key target for anti-plague therapy. However, little information is available about the molecular aspects of its function and interactions with the human host, and the structure of Ail is not known. Here we describe the recombinant expression, purification, refolding, and sample preparation of Ail for solution and solid-state NMR structural studies in lipid micelles and lipid bilayers. The initial NMR and CD spectra show that Ail adopts a well-defined transmembrane β-sheet conformation in lipids.
authors.affiliation: Sanford Burnham Medical Research Institute, 10901 North Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Leigh A
authors.initials: LA
authors.lastname: Plesniak
authors.forename: Radhakrishnan
authors.initials: R
authors.lastname: Mahalakshmi
authors.forename: Candace
authors.initials: C
authors.lastname: Rypien
authors.forename: Yuan
authors.initials: Y
authors.lastname: Yang
authors.forename: Jasmina
authors.initials: J
authors.lastname: Racic
authors.forename: Francesca M
authors.initials: FM
authors.lastname: Marassi
mesh_terms: D000595:Amino Acid Sequence; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D002942:Circular Dichroism; D015964:Gene Expression Regulation, Bacterial; D006863:Hydrogen-Ion Concentration; D008055:Lipids / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017386:Sequence Homology, Amino Acid; D037521:Virulence Factors / Q000737:chemistry* / Q000378:metabolism; D015010:Yersinia pestis / Q000378:metabolism*
chemical_list: C526543:Ail protein, Yersinia pestis; D001425:Bacterial Outer Membrane Proteins; D008055:Lipids; D037521:Virulence Factors