BhCRASP-1 of the relapsing fever spirochete Borrelia hermsii is a factor H and plasminogen binding protein

2007 ◽  
Vol 44 (16) ◽  
pp. 3986
Author(s):  
Michael Kirschfink ◽  
Evelyn Rossmann ◽  
Pia Herzberger ◽  
Christine Skerka ◽  
Peter F. Zipfel ◽  
...  
Keyword(s):  
Binding Protein ◽  
Factor H ◽  
Relapsing Fever ◽  
Borrelia Hermsii

BhCRASP-1 of the relapsing fever spirochete Borrelia hermsii is a factor H- and plasminogen-binding protein

2008 ◽  
Vol 298 ◽  
pp. 272-283 ◽  
Author(s):  
Evelyn Rossmann ◽  
Peter Kraiczy ◽  
Pia Herzberger ◽  
Christine Skerka ◽  
Michael Kirschfink ◽  
...  
Keyword(s):  
Binding Protein ◽  
Factor H ◽  
Relapsing Fever ◽  
Borrelia Hermsii

scholarly journals Immunological and Molecular Analyses of the Borrelia hermsii Factor H and Factor H-Like Protein 1 Binding Protein, FhbA: Demonstration of Its Utility as a Diagnostic Marker and Epidemiological Tool for Tick-Borne Relapsing Fever

2006 ◽  
Vol 74 (8) ◽  
pp. 4519-4529 ◽  
Author(s):  
Kelley M. Hovis ◽  
Martin E. Schriefer ◽  
Sonia Bahlani ◽  
Richard T. Marconi
Keyword(s):  
Binding Protein ◽  
Pcr Primers ◽  
Factor H ◽  
Antigenic Structure ◽  
Diagnostic Tools ◽  
Relapsing Fever ◽  
Borrelia Hermsii ◽  
Specific Pcr ◽  
Epidemiological Tool ◽  
Species Specific

ABSTRACT It has been demonstrated that Borrelia hermsii, a causative agent of relapsing fever, produces a factor H (FH) and FH-like protein 1 (FHL-1) binding protein. The binding protein has been designated FhbA. To determine if FH/FHL-1 binding is widespread among B. hermsii isolates, a diverse panel of strains was tested for the FH/FHL-1 binding phenotype and FhbA production. Most isolates (23/24) produced FhbA and bound FH/FHL-1. Potential variation in FhbA among isolates was analyzed by DNA sequence analyses. Two genetically distinct FhbA types, designated fhbA1 and fhbA2, were delineated, and type-specific PCR primers were generated to allow for rapid differentiation. Pulsed-field gel electrophoresis and hybridization analyses demonstrated that all isolates that possess the gene carry it on a 200-kb linear plasmid (lp200), whereas isolates that lack the gene lack lp200 and instead carry an lp170. To determine if FhbA is antigenic during infection and to assess the specificity of the response, recombinant FhbA1 (rFhbA1) and rFhbA2 were screened with serum from infected mice and humans. FhbA was found to be expressed and antigenic and to elicit a potentially type-specific FhbA response. To localize the epitopes of FhbA1 and FhbA2, truncations were generated and screened with infection serum. The epitopes were determined to be conformationally defined. Collectively, these analyses indicate that FH/FHL-1 binding is a widespread virulence mechanism for B. hermsii and provide insight into the genetic and antigenic structure of FhbA. The data also have potential implications for understanding the epidemiology of relapsing fever in North America and can be applied to the future development of species-specific diagnostic tools.

scholarly journals Identification and Characterization of a Linear-Plasmid-Encoded Factor H-Binding Protein (FhbA) of the Relapsing Fever Spirochete Borrelia hermsii

2004 ◽  
Vol 186 (9) ◽  
pp. 2612-2618 ◽  
Author(s):  
Kelley M. Hovis ◽  
John V. McDowell ◽  
LaToya Griffin ◽  
Richard T. Marconi
Keyword(s):  
Molecular Mechanisms ◽  
Binding Protein ◽  
Genetic Locus ◽  
Factor H ◽  
Linear Plasmid ◽  
Relapsing Fever ◽  
Gene Encoding ◽  
Borrelia Hermsii ◽  
Identification And Characterization

ABSTRACT In North America, tick-borne relapsing fever (TBRF) is caused by the spirochete species Borrelia hermsii, Borrelia parkeri, and Borrelia turicatae. We previously demonstrated that some isolates of B. hermsii and B. parkeri are capable of binding factor H and that cell-bound factor H can participate in the factor I-mediated cleavage of C3b. Isolates that bound factor H expressed a factor H-binding protein (FHBP) that we estimated to be approximately 19 to 20 kDa in size and thus, pending further characterization, temporarily designated FHBP19. Until this report, none of the FHBPs of the TBRF spirochetes had been characterized. Here we have recovered the gene encoding the FHBP of B. hermsii YOR from a lambda ZAP II library and determined its sequence. The gene encodes a full-length protein of 22.7 kDa, which after processing is predicted to be 20.5 kDa. This protein, which we redesignate factor H-binding protein A (FhbA), is unique to B. hermsii. Two-dimensional pulsed-field gel electrophoresis and hybridization analyses revealed that the B. hermsii gene encoding FhbA is a single genetic locus that maps to a linear plasmid of approximately 220 kb. The general properties of FhbA were also assessed. The protein was found to be surface exposed and lipidated. Analysis of the antibody response to FhbA in infected mice revealed that it is antigenic during infection, indicating expression during infection. The identification and characterization of FhbA provides further insight into the molecular mechanisms of pathogenesis of the relapsing fever spirochetes.

scholarly journals The Borrelia hermsii Factor H Binding Protein FhbA Is Not Required for Infectivity in Mice or for Resistance to Human ComplementIn Vitro

2014 ◽  
Vol 82 (8) ◽  
pp. 3324-3332 ◽  
Author(s):  
Lindy M. Fine ◽  
Daniel P. Miller ◽  
Katherine L. Mallory ◽  
Brittney K. Tegels ◽  
Christopher G. Earnhart ◽  
...  
Keyword(s):  
Genetic Manipulation ◽  
Binding Protein ◽  
Factor H ◽  
Negative Regulator ◽  
Human Complement ◽  
Relapsing Fever ◽  
Borrelia Hermsii ◽  
Content Type

ABSTRACTThe primary causative agent of tick-borne relapsing fever in North America isBorrelia hermsii. It has been hypothesized thatB. hermsiievades complement-mediated destruction by binding factor H (FH), a host-derived negative regulator of complement.In vitro,B. hermsiiproduces a single FH binding protein designated FhbA (FH binding protein A). The properties and ligand binding activity of FhbA suggest that it plays multiple roles in pathogenesis. It binds plasminogen and has been identified as a significant target of a B1b B cell-mediated IgM response in mice. FhbA has also been explored as a potential diagnostic antigen forB. hermsiiinfection in humans. The ability to test the hypothesis that FhbA is a critical virulence factorin vivohas been hampered by the lack of well-developed systems for the genetic manipulation of the relapsing fever spirochetes. In this report, we have successfully generated aB. hermsiifhbAdeletion mutant (theB. hermsiiYORΔfhbAstrain) through allelic exchange mutagenesis. Deletion offhbAabolished FH binding by the YORΔfhbAstrain and eliminated cleavage of C3b on the cell surface. However, the YORΔfhbAstrain remained infectious in mice and retained resistance to killingin vitroby human complement. Collectively, these results indicate thatB. hermsiiemploys an FhbA/FH-independent mechanism of complement evasion that allows for resistance to killing by human complement and persistence in mice.

scholarly journals Fibronectin-Binding Protein of Borrelia hermsii Expressed in the Blood of Mice with Relapsing Fever

2014 ◽  
Vol 82 (6) ◽  
pp. 2520-2531 ◽  
Author(s):  
E. R. G. Lewis ◽  
R. A. Marcsisin ◽  
S. A. Campeau Miller ◽  
F. Hue ◽  
A. Phillips ◽  
...  
Keyword(s):  
Binding Protein ◽  
Relapsing Fever ◽  
Borrelia Hermsii ◽  
Fibronectin Binding Protein ◽  
Fibronectin Binding

scholarly journals Relapsing Fever Spirochetes Borrelia recurrentis and B. duttonii Acquire Complement Regulators C4b-Binding Protein and Factor H

2006 ◽  
Vol 74 (7) ◽  
pp. 4157-4163 ◽  
Author(s):  
T. Meri ◽  
S. J. Cutler ◽  
A. M. Blom ◽  
S. Meri ◽  
T. S. Jokiranta
Keyword(s):  
Binding Protein ◽  
Protein A ◽  
Sensu Stricto ◽  
Factor H ◽  
Complement Pathway ◽  
Relapsing Fever ◽  
Borrelia Recurrentis ◽  
First Report ◽  
Complement Regulators

ABSTRACT Relapsing fever is a rapidly progressive and severe septic disease caused by certain Borrelia spirochetes. The disease is divided into two forms, i.e., epidemic relapsing fever, caused by Borrelia recurrentis and transmitted by lice, and the endemic form, caused by several Borrelia species, such as B. duttonii, and transmitted by soft-bodied ticks. The spirochetes enter the bloodstream by the vector bite and live persistently in plasma even after the development of specific antibodies. This leads to fever relapses and high mortality and clearly indicates that the Borrelia organisms utilize effective immune evasion strategies. In this study, we show that the epidemic relapsing fever pathogen B. recurrentis and an endemic relapsing fever pathogen, B. duttonii, are serum resistant, i.e., resistant to complement in vitro. They acquire the host alternative complement pathway regulator factor H on their surfaces in a similar way to that of the less serum-resistant Lyme disease pathogen, B. burgdorferi sensu stricto. More importantly, the relapsing fever spirochetes specifically bind host C4b-binding protein, a major regulator of the antibody-mediated classical complement pathway. Both complement regulators retained their functional activities when bound to the surfaces of the spirochetes. In conclusion, this is the first report of complement evasion by Borrelia recurrentis and B. duttonii and the first report showing capture of C4b-binding protein by spirochetes.

Molecular and immunological analyses of the Borrelia hermsii factor H/FHL-1 binding protein, FhbA

2007 ◽  
Vol 44 (1-3) ◽  
pp. 185
Author(s):  
Kelley M. Hovis ◽  
Martin E. Schriefer ◽  
Tania Sadlon ◽  
David L. Gordon ◽  
Richard T. Marconi
Keyword(s):  
Binding Protein ◽  
Factor H ◽  
Borrelia Hermsii

scholarly journals Molecular Analyses of the Interaction of Borrelia hermsii FhbA with the Complement Regulatory Proteins Factor H and Factor H-Like Protein 1

2006 ◽  
Vol 74 (4) ◽  
pp. 2007-2014 ◽  
Author(s):  
Kelley M. Hovis ◽  
Janice P. Jones ◽  
Tania Sadlon ◽  
Gauri Raval ◽  
David L. Gordon ◽  
...  
Keyword(s):  
Contact Point ◽  
Regulatory Protein ◽  
Protein A ◽  
Random Mutagenesis ◽  
Binding Pocket ◽  
Factor H ◽  
Intramolecular Interactions ◽  
Relapsing Fever ◽  
Borrelia Hermsii ◽  
Loop Region

ABSTRACT Borrelia hermsii, the primary etiological agent of tick-borne relapsing fever in North America, binds the complement regulatory protein factor H (FH) as a means of evading opsonophagocytosis and the alternative complement pathway. The ability of FH-binding protein A (FhbA) to bind FH-like protein 1 (FHL-1) has not been assessed previously. In this study, using a whole-cell absorption assay, we demonstrated that B. hermsii absorbs both FH and FHL-1 from human serum. Consistent with this, affinity ligand binding immunoblot analyses revealed that FH constructs spanning short consensus repeats 1 to 7 and 16 to 20 bind to FhbA. To investigate the molecular basis of the interaction of FhbA with FH/FHL-1, recombinant FhbA truncated proteins were generated and tested for FH/FHL-1 binding. Binding required determinants located in both the N- and C-terminal domains of FhbA, suggesting that long-range intramolecular interactions are involved in the formation and presentation of the FH/FHL-1-binding pocket. To identify specific FhbA residues involved in binding, random mutagenesis was performed. These analyses identified a loop region of FhbA that may serve as a contact point for FH/FHL-1. The data presented here expand our understanding of the pathogenic mechanisms of the relapsing fever spirochetes and of the molecular nature of the interaction between FH/FHL-1 and FhbA.

scholarly journals Identification of an Antiparallel Coiled-Coil/Loop Domain Required for Ligand Binding by the Borrelia hermsii FhbA Protein: Additional Evidence for the Role of FhbA in the Host-Pathogen Interaction

2008 ◽  
Vol 76 (5) ◽  
pp. 2113-2122 ◽  
Author(s):  
Kelley M. Hovis ◽  
John C. Freedman ◽  
Hongming Zhang ◽  
Jonathan L. Forbes ◽  
Richard T. Marconi
Keyword(s):  
Serum Proteins ◽  
Coiled Coil ◽  
Factor H ◽  
Site Directed Mutagenesis ◽  
Relapsing Fever ◽  
Borrelia Hermsii ◽  
Alpha Helices ◽  
Host Pathogen Interaction ◽  
Loop Domain ◽  
Host Pathogen

ABSTRACT Borrelia hermsii, an etiological agent of tick-borne relapsing fever in North America, binds host-derived serum proteins including factor H (FH), plasminogen, and an unidentified 60-kDa protein via its FhbA protein. Two distinct phylogenetic types of FhbA have been delineated (FhbA1 and FhbA2). These orthologs share a conserved C-terminal domain that contains two alpha helices with a high predictive probability of coiled-coil formation that are separated by a 14-amino-acid loop domain. Through site-directed mutagenesis, we have identified residues within these domains that influence the binding of both mouse and human FH, plasminogen, and/or the 60-kDa protein. To further investigate the involvement of FhbA in the host-pathogen interaction, strains that are either FhbA+ (isolate YOR) or FhbA− (isolate REN) were tested for serum sensitivity. Significant differences were observed, with YOR and REN being serum resistant and serum sensitive (intermediate), respectively. To test the abilities of these strains to infect and persist in mice, mice were needle inoculated, and infectivity and persistence were then assessed. While both strains REN and YOR infected mice, only the FhbA+ YOR strain persisted beyond day 4. Survival of the YOR isolate in blood correlated with the upregulation of the fhbA gene, as demonstrated by real-time reverse transcriptase PCR. These data advance our understanding of the unique interactions of FhbA with individual serum proteins and provide support for the hypothesis that FhbA is an important contributor to the pathogenesis of the relapsing fever spirochete B. hermsii.

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