title: Structure and biochemical analysis of a secretin pilot protein.
abstract: The ability to translocate virulence proteins into host cells through a type III secretion apparatus (TTSS) is a hallmark of several Gram-negative pathogens including Shigella, Salmonella, Yersinia, Pseudomonas, and enteropathogenic Escherichia coli. In common with other types of bacterial secretion apparatus, the assembly of the TTSS complex requires the preceding formation of its integral outer membrane secretin ring component. We have determined at 1.5 A the structure of MxiM28-142, the Shigella pilot protein that is essential for the assembly and membrane association of the Shigella secretin, MxiD. This represents the first atomic structure of a secretin pilot protein from the several bacterial secretion systems containing an orthologous secretin component. A deep hydrophobic cavity is observed in the novel 'cracked barrel' structure of MxiM, providing a specific binding domain for the acyl chains of bacterial lipids, a proposal that is supported by our various lipid/MxiM complex structures. Isothermal titration analysis shows that the C-terminal domain of the secretin, MxiD525-570, hinders lipid binding to MxiM.
authors.affiliation: Department of Biochemistry, University of British Columbia, Vancouver, BC, Canada.
authors.forename: Paula I
authors.initials: PI
authors.lastname: Lario
authors.forename: Richard A
authors.initials: RA
authors.lastname: Pfuetzner
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Frey
authors.forename: Louise
authors.initials: L
authors.lastname: Creagh
authors.forename: Charles
authors.initials: C
authors.lastname: Haynes
authors.forename: Anthony T
authors.initials: AT
authors.lastname: Maurelli
authors.forename: Natalie C J
authors.initials: NC
authors.lastname: Strynadka
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000378:metabolism* / Q000502:physiology; D003460:Crystallization; D008074:Lipoproteins / Q000737:chemistry* / Q000502:physiology; D011487:Protein Conformation; D012763:Shigella flexneri / Q000502:physiology*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D008074:Lipoproteins; C079723:MxiD protein, Shigella; C078384:mxiM protein, Shigella flexneri