title: Extracellular secretion of the Borrelia burgdorferi Oms28 porin and Bgp, a glycosaminoglycan binding protein.
abstract: Borrelia burgdorferi, the Lyme disease pathogen, cycles between its Ixodes tick vector and vertebrate hosts, adapting to vastly different biochemical environments. Spirochete gene expression as a function of temperature, pH, growth phase, and host milieu is well studied, and recent work suggests that regulatory networks are involved. Here, we examine the release of Borrelia burgdorferi strain B31 proteins into conditioned medium. Spirochetes intrinsically radiolabeled at concentrations ranging from 10(7) to 10(9) cells per ml secreted Oms28, a previously characterized outer membrane porin, into RPMI medium. As determined by immunoblotting, this secretion was not associated with outer membrane blebs or cytoplasmic contamination. A similar profile of secreted proteins was obtained for spirochetes radiolabeled in mixtures of RPMI medium and serum-free Barbour-Stoenner-Kelly (BSK II) medium. Proteomic liquid chromatography-tandem mass spectrometry analysis of tryptic fragments derived from strain B31 culture supernatants confirmed the identity of the 28-kDa species as Oms28 and revealed a 26-kDa protein as 5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase (Pfs-2), previously described as Bgp, a glycosaminoglycan-binding protein. The release of Oms28 into the culture medium is more selective when the spirochetes are in logarithmic phase of growth compared to organisms obtained from stationary phase. As determined by immunoblotting, stationary-phase spirochetes released OspA, OspB, and flagellin. Oms28 secreted by strains B31, HB19, and N40 was also recovered by radioimmunoprecipitation. This is the first report of B. burgdorferi protein secretion into the extracellular environment. The possible roles of Oms28 and Bgp in the host-pathogen interaction are considered.
authors.affiliation: Department of Chemistry and Biochemistry, Middlebury College, VT 05753, USA. rcluss@middlebury.edu
authors.forename: Robert G
authors.initials: RG
authors.lastname: Cluss
authors.forename: Damon A
authors.initials: DA
authors.lastname: Silverman
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Stafford
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D025065:Borrelia burgdorferi / Q000254:growth & development* / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism*; D017077:Culture Media, Conditioned / Q000737:chemistry; D015964:Gene Expression Regulation, Bacterial; D047908:Intracellular Signaling Peptides and Proteins; D018272:Porins / Q000378:metabolism*; D015531:Radioimmunoprecipitation Assay
chemical_list: D001426:Bacterial Proteins; C474476:Bgp protein, Borrelia burgdorferi; D002352:Carrier Proteins; D017077:Culture Media, Conditioned; D047908:Intracellular Signaling Peptides and Proteins; D018272:Porins; C100895:protein Oms28, Borrelia burgdorferi