title: Species-specific functioning of the Pseudomonas XcpQ secretin: role for the C-terminal homology domain and lipopolysaccharide.
abstract: Secretins are oligomeric proteins that mediate the export of macromolecules across the bacterial outer membrane. The members of the secretin superfamily possess a C-terminal homology domain that is important for oligomerization and channel formation, while their N-terminal halves are thought to be involved in system-specific interactions. The XcpQ secretin of Pseudomonas spp. is a component of the type II secretion pathway. XcpQ from Pseudomonas alcaligenes is not able to functionally replace the secretin of the closely related species Pseudomonas aeruginosa. By analysis of chimeric XcpQ proteins, a region important for species-specific functioning was mapped between amino acid residues 344 and 478 in the C-terminal homology domain. Two chromosomal suppressor mutations were obtained that resulted in the proper functioning in P. aeruginosa of P. alcaligenes XcpQ and inactive hybrids. These mutations caused a defect in the synthesis of the lipopolysaccharide (LPS) outer core region. Subsequent analysis of different LPS mutants showed that changes in the outer core and not the loss of O antigen caused the suppressor phenotype. High concentrations of divalent cations in the growth medium also allowed P. alcaligenes XcpQ and inactive hybrids to function properly in P. aeruginosa. Since divalent cations are known to affect the structure of LPS, this observation supports the hypothesis that LPS has a role in the functioning of secretins.
authors.affiliation: Department of Molecular Microbiology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Wilbert
authors.initials: W
authors.lastname: Bitter
authors.forename: Ria
authors.initials: R
authors.lastname: van Boxtel
authors.forename: Mathijs
authors.initials: M
authors.lastname: Groeneweg
authors.forename: Patricia Sánchez
authors.initials: PS
authors.lastname: Carballo
authors.forename: Ulrich
authors.initials: U
authors.lastname: Zähringer
authors.forename: Jan
authors.initials: J
authors.lastname: Tommassen
authors.forename: Margot
authors.initials: M
authors.lastname: Koster
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism*; D002413:Cations, Divalent; D008070:Lipopolysaccharides; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism*; D017434:Protein Structure, Tertiary / Q000502:physiology; D011549:Pseudomonas / Q000737:chemistry / Q000378:metabolism*; D013045:Species Specificity; D013329:Structure-Activity Relationship
chemical_list: D001426:Bacterial Proteins; D002413:Cations, Divalent; D008070:Lipopolysaccharides; D008565:Membrane Proteins