title: Dynamics of Klebsiella pneumoniae OmpA transmembrane domain: the four extracellular loops display restricted motion behavior in micelles and in lipid bilayers.
abstract: The transmembrane domain of Klebsiella pneumoniae OmpA (KpOmpA) possesses four long extracellular loops that exhibit substantial sequence variability throughout OmpA homologs in Enterobacteria, in comparison with the highly conserved membrane-embedded β-barrel core. These loops are responsible for the immunological properties of the protein, including cellular and humoral recognition. In addition to key features revealed by structural elucidation of the KpOmpA transmembrane domain in detergent micelles, studies of protein dynamics provide insight into its function and/or mechanism of action. We have investigated the dynamics of KpOmpA in a lipid bilayer, using magic angle spinning solid-state NMR. The dynamics of the β-barrel and loop regions were probed by the spin-lattice relaxation times of the C(α) and C(β) atoms of the serine and threonine residues, and by cross-polarization dynamics. The β-barrel core of the protein is rigid; the C-terminal halves of two of the four extracellular loops (L1 and L3), which are particularly long in KpOmpA, are highly mobile. The other two loops (L2 and L4), which are very similar to their homologs in Escherichia coli OmpA, and the N-terminal halves of L1 and L3 exhibit more restricted motions. We suggest a correlation between the sequence variability and the dynamics of certain loop regions, which accounts for their respective contributions to the structural and immunological properties of the protein.
authors.affiliation: Institute of Pharmacology and Structural Biology, Université de Toulouse, Toulouse, France. iordan.iordanov@ipbs.fr
authors.forename: Iordan
authors.initials: I
authors.lastname: Iordanov
authors.forename: Marie
authors.initials: M
authors.lastname: Renault
authors.forename: Valérie
authors.initials: V
authors.lastname: Réat
authors.forename: Patrick D
authors.initials: PD
authors.lastname: Bosshart
authors.forename: Andreas
authors.initials: A
authors.lastname: Engel
authors.forename: Olivier
authors.initials: O
authors.lastname: Saurel
authors.forename: Alain
authors.initials: A
authors.lastname: Milon
mesh_terms: D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000502:physiology*; D002499:Centrifugation, Density Gradient; D003902:Detergents / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D007711:Klebsiella pneumoniae / Q000378:metabolism*; D008051:Lipid Bilayers / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008823:Micelles; D046529:Microscopy, Electron, Transmission / Q000379:methods; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013395:Sucrose / Q000737:chemistry
chemical_list: D001425:Bacterial Outer Membrane Proteins; D003902:Detergents; D008051:Lipid Bilayers; D008823:Micelles; C078167:OMPA outer membrane proteins; D013395:Sucrose