Mutational analyses of the BbCRASP-1 protein of Borrelia burgdorferi identify residues relevant for the architecture and binding of host complement regulators FHL-1 and factor H

Kraiczy, Peter; Hanssen-Hübner, Christa; Kitiratschky, Veronique; Brenner, Christiane; Besier, Silke; Brade, Volker; Simon, Markus M.; Skerka, Christine; Roversi, Pietro; Lea, Susan M.; Stevenson, Brian; Wallich, Reinhard; Zipfel, Peter F.

doi:10.1016/j.ijmm.2008.09.002

Cited by 24 publications

(28 citation statements)

References 72 publications

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“…The same membrane was also immunoblotted with rabbit anti-FlaB antibodies to ensure equal loading between all samples. (15,16,25,35,38,41,67,67,67,71). Consistent with these prior observations, we previously generated a CspA mutant in an avirulent strain of B. burgdorferi and reported that CspA confers resistance to human serum in vitro (10).…”

Section: Methodssupporting

confidence: 81%

CspA-Mediated Binding of Human Factor H Inhibits Complement Deposition and Confers Serum Resistance in Borrelia burgdorferi

et al. 2009

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Borrelia burgdorferi has developed efficient mechanisms for evading the innate immune response during mammalian infection and has been shown to be resistant to the complement-mediated bactericidal activity of human serum. It is well recognized that B. burgdorferi expresses multiple lipoproteins on its surface that bind the human complement inhibitors factor H and factor H-like protein 1 (FH/FHL-1). The binding of FH/FHL-1 on the surface of B. burgdorferi is thought to enhance its ability to evade serum-mediated killing during the acute phase of infection. One of the key B. burgdorferi FH/FHL-1 binding proteins identified thus far was designated CspA. While it is known that CspA binds FH/FHL-1, it is unclear how the interaction between CspA and FH/FHL-1 specifically enhances serum resistance. To better understand how CspA mediates serum resistance in B. burgdorferi, we inactivated cspA in a virulent strain of B. burgdorferi. An affinity ligand blot immunoassay and indirect immunofluorescence revealed that the CspA mutant does not efficiently bind human FH to its surface. Consistent with the lack of FH binding, the CspA mutant was also highly sensitive to killing by human serum. Additionally, the deposition of complement components C3, C6, and C5b-9 was enhanced on the surface of the CspA mutant compared to that of the wild-type strain. The combined data lead us to conclude that the CspA-mediated binding of human FH confers serum resistance by directly inhibiting complement deposition on the surface of B. burgdorferi.

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Section: Methodssupporting

confidence: 81%

CspA-Mediated Binding of Human Factor H Inhibits Complement Deposition and Confers Serum Resistance in Borrelia burgdorferi

et al. 2009

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“…To date, majority of the studies are focused on the interaction between human factor H and FHBPs of Borrelia Hitherto, binding site for human fH on BbCRASP-1 molecule has been described [23,24], however, scanty reports are available, if at all, which show binding pockets for FHBPs present on fH protein from various hosts. Study of comparative binding ability of FHBPs to various domains of fH (like sushi 6-7 and sushi [19][20], depending on the host species, might be the promising way to understand fundaments of FHBPs:fH interaction.…”

Section: Discussionmentioning

confidence: 99%

Variable regions in the sushi domains 6–7 and 19–20 of factor H in animals and human lead to change in the affinity to factor H binding protein of Borrelia

Bhide

Pulzová

et al. 2012

Journal of Proteomics

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“…Construction and expression of plasmids harboring the cspA, cspZ, erpP, erpC, and erpA genes encoding CspA, CspZ, ErpP, ErpC, and ErpA, respectively, and purification of the proteins were described (13,18,37,38). Cloning and expression of the CspA deletion mutants were performed as previously described (18,39). For generation of the CspA deletion mutants, specific primers (Table 1) were used, and the inserts were recloned into the pQE-30 Xa vector (Qiagen).…”

Section: Methodsmentioning

confidence: 99%

CspA from Borrelia burgdorferi inhibits the terminal complement pathway

Hallström¹,

Siegel²,

Mörgelin³

et al. 2013

Molecular Immunology

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In order to survive and persist in an immunocompetent human host, Borrelia burgdorferi controls the human immune attack and blocks the damaging effects of the activated complement system. These Gram-negative spirochetes use CspA (CRASP-1) and four additional immune evasion proteins to bind combinations of human plasma regulators, including factor H, factor H-like protein 1 (FHL-1), complement factor H-related protein 1 (CFHR1), CFHR2, CFHR5, and plasminogen. As many microbial immune evasion proteins have multiple functions, we hypothesized that CspA has additional roles in complement or immune control. Here, we identify CspA as a terminal complement inhibitor. Borrelial CspA binds the human terminal complement components C7 and C9 and blocks assembly and membrane insertion of the terminal complement complex (TCC). CspA inhibits TCC assembly at the level of C7, as revealed by hemolytic assays, and inhibits polymerization of C9. CspA, when ectopically expressed on the surface of serum-sensitive Borrelia garinii, blocks TCC assembly on the level of C7 and induces serum resistance in the transformed bacteria. This CspA-mediated serum resistance and terminal complement pathway inhibition allow B. burgdorferi to survive in the hostile environment of human plasma.

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Mutational analyses of the BbCRASP-1 protein of Borrelia burgdorferi identify residues relevant for the architecture and binding of host complement regulators FHL-1 and factor H

Cited by 24 publications

References 72 publications

CspA-Mediated Binding of Human Factor H Inhibits Complement Deposition and Confers Serum Resistance in Borrelia burgdorferi

CspA-Mediated Binding of Human Factor H Inhibits Complement Deposition and Confers Serum Resistance in Borrelia burgdorferi

Variable regions in the sushi domains 6–7 and 19–20 of factor H in animals and human lead to change in the affinity to factor H binding protein of Borrelia

CspA from Borrelia burgdorferi inhibits the terminal complement pathway

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