scholarly journals Structural characterization of CspZ, a complement regulator factor H and FHL-1 binding protein fromBorrelia burgdorferi

FEBS Journal ◽  
2014 ◽  
Vol 281 (11) ◽  
pp. 2613-2622 ◽  
Author(s):  
Kalvis Brangulis ◽  
Ivars Petrovskis ◽  
Andris Kazaks ◽  
Janis Bogans ◽  
Martins Otikovs ◽  
...  
Keyword(s):  
Structural Characterization ◽  
Binding Protein ◽  
Factor H ◽  
Complement Regulator

scholarly journals Identification and Functional Characterization of Complement Regulator-Acquiring Surface Protein 1 of the Lyme Disease Spirochetes Borrelia afzelii and Borrelia garinii

2005 ◽  
Vol 73 (4) ◽  
pp. 2351-2359 ◽  
Author(s):  
Reinhard Wallich ◽  
Joseph Pattathu ◽  
Veronique Kitiratschky ◽  
Christiane Brenner ◽  
Peter F. Zipfel ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Binding Site ◽  
Binding Protein ◽  
Functional Characterization ◽  
Surface Protein ◽  
Factor H ◽  
Borrelia Garinii ◽  
Borrelia Afzelii ◽  
Complement Regulator

ABSTRACT Complement regulator-acquiring surface protein 1 (CRASP-1) is the dominant factor-H-like protein 1 (FHL-1)- and factor-H-binding protein of Borrelia burgdorferi and is suggested to contribute to persistence of the pathogen. The prototype CRASP-1 of B. burgdorferi sensu stricto (CRASP-1Bb) has been formerly characterized. As shown recently, serum-resistant Borrelia afzelii strains express a unique FHL-1 and factor H-binding protein, designated CRASP-1Ba. Here, we describe for the first time the isolation and functional characterization of the gene encoding the full-length CRASP-1Ba of 28 kDa, which, upon processing, is predicted to be 26.4 kDa. CPASP-1Ba of B. afzelii spirochetes is associated with a genetic locus encoding the orthologous gbb54 gene family that maps to the linear plasmid of approximately 54 kb. Ligand affinity blotting techniques demonstrate that both native and recombinant CRASP-1Ba molecules strongly bind to FHL-1 and much more weakly to factor H. The FHL-1 and factor-H-binding site in CRASP-1Ba is shown to be localized to a 12-amino-acid residue domain at the C terminus of the protein. For comparison, the corresponding cspA-like gene(s) of a serum-sensitive Borrelia garinii strain has also been cloned and characterized. Most notably, two CRASP-1-related B. garinii proteins were identified; however, both molecules bind only weakly to FHL-1 and not at all to factor H. The present identification of the binding site of CRASP-1Ba represents an important step forward in our understanding of the pathogenesis of Lyme disease and may be helpful to design therapeutic regimens to interfere with complement evasion strategies of human pathogenic Borrelia strains.

Structural Characterization of HP1264 Reveals a Novel Fold for the Flavin Mononucleotide Binding Protein

Biochemistry ◽  
2013 ◽  
Vol 52 (9) ◽  
pp. 1583-1593 ◽  
Author(s):  
Ki-Young Lee ◽  
Ji-Hun Kim ◽  
Kyu-Yeon Lee ◽  
Jiyun Lee ◽  
Ingyun Lee ◽  
...  
Keyword(s):  
Structural Characterization ◽  
Binding Protein ◽  
Flavin Mononucleotide

scholarly journals Characterization of Diverse Subvariants of the Meningococcal Factor H (fH) Binding Protein for Their Ability To Bind fH, To Mediate Serum Resistance, and To Induce Bactericidal Antibodies

2010 ◽  
Vol 79 (2) ◽  
pp. 970-981 ◽  
Author(s):  
Kate L. Seib ◽  
Brunella Brunelli ◽  
Barbara Brogioni ◽  
Emmanuelle Palumbo ◽  
Stefania Bambini ◽  
...  
Keyword(s):  
Binding Protein ◽  
Genetically Engineered ◽  
Factor H ◽  
Serum Resistance ◽  
Dependent Manner ◽  
Bacterial Surface ◽  
Bactericidal Antibody ◽  
Human Sera ◽  
The Stability

ABSTRACTNeisseria meningitidisis a commensal of the human nasopharynx but is also a major cause of septicemia and meningitis. The meningococcal factor H binding protein (fHbp) binds human factor H (fH), enabling downregulation of complement activation on the bacterial surface. fHbp is a component of two serogroup B meningococcal vaccines currently in clinical development. Here we characterize 12 fHbp subvariants for their level of surface exposure and ability to bind fH, to mediate serum resistance, and to induce bactericidal antibodies. Flow cytometry and Western analysis revealed that all strains examined expressed fHbp on their surface to different extents and bound fH in an fHbp-dependent manner. However, differences in fH binding did not always correlate with the level of fHbp expression, indicating that this is not the only factor affecting the amount of fH bound. To overcome the issue of strain variability in fHbp expression, the MC58ΔfHbpstrain was genetically engineered to express different subvariants from a constitutive heterologous promoter. These recombinant strains were characterized for fH binding, and the data confirmed that each subvariant binds different levels of fH. Surface plasmon resonance revealed differences in the stability of the fHbp-fH complexes that ranged over 2 orders of magnitude, indicating that differences in residues between and within variant groups can influence fH binding. Interestingly, the level of survival in human sera of recombinant MC58 strains expressing diverse subvariants did not correlate with the level of fH binding, suggesting that the interaction of fHbp with fH is not the only function of fHbp that influences serum resistance. Furthermore, cross-reactive bactericidal activity was seen within each variant group, although the degree of activity varied, suggesting that amino acid differences within each variant group influence the bactericidal antibody response.

Insulin-like growth factor binding protein-2: NMR analysis and structural characterization of the N-terminal domain

Biochimie ◽  
2012 ◽  
Vol 94 (3) ◽  
pp. 608-616 ◽  
Author(s):  
Charles A. Galea ◽  
Mehdi Mobli ◽  
Kerrie A. McNeil ◽  
Terrence D. Mulhern ◽  
John C. Wallace ◽  
...  
Keyword(s):  
Growth Factor ◽  
Structural Characterization ◽  
Binding Protein ◽  
Insulin Like Growth Factor ◽  
Nmr Analysis ◽  
Factor Binding ◽  
Terminal Domain

scholarly journals Structural Characterization of the Fibroblast Growth Factor-binding Protein Purified from Bovine Prepartum Mammary Gland Secretion

2000 ◽  
Vol 275 (26) ◽  
pp. 19469-19474 ◽  
Author(s):  
Rene Lametsch ◽  
Jan T. Rasmussen ◽  
Laust B. Johnsen ◽  
Stig Purup ◽  
Kristen Sejrsen ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Structural Characterization ◽  
Binding Protein ◽  
Gland Secretion ◽  
Fibroblast Growth ◽  
Factor Binding

scholarly journals Characterization of Neisseria meningitidis Isolates That Do Not Express the Virulence Factor and Vaccine Antigen Factor H Binding Protein

2011 ◽  
Vol 18 (6) ◽  
pp. 1002-1014 ◽  
Author(s):  
Jay Lucidarme ◽  
Lionel Tan ◽  
Rachel M. Exley ◽  
Jamie Findlow ◽  
Ray Borrow ◽  
...  
Keyword(s):  
Neisseria Meningitidis ◽  
Binding Protein ◽  
Factor H ◽  
Negative Regulator ◽  
Vaccine Antigen ◽  
Natural Immunity ◽  
Content Type ◽  
Reading Frame ◽  
Flanking Sequences

ABSTRACTNeisseria meningitidisremains a leading cause of bacterial sepsis and meningitis. Complement is a key component of natural immunity against this important human pathogen, which has evolved multiple mechanisms to evade complement-mediated lysis. One approach adopted by the meningococcus is to recruit a human negative regulator of the complement system, factor H (fH), to its surface via a lipoprotein, factor H binding protein (fHbp). Additionally, fHbp is a key antigen in vaccines currently being evaluated in clinical trials. Here we characterize strains ofN. meningitidisfrom several distinct clonal complexes which do not express fHbp; all strains were recovered from patients with disseminated meningococcal disease. We demonstrate that these strains have either a frameshift mutation in thefHbpopen reading frame or have entirely lostfHbpand some flanking sequences. No fH binding was detected to other ligands among thefHbp-negative strains. The implications of these findings for meningococcal pathogenesis and prevention are discussed.

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.

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