EurekaMag.com logo
+ Translate

Identification of an immunologically important hypervariable domain of major outer surface protein A of Borrelia burgdorferi


, : Identification of an immunologically important hypervariable domain of major outer surface protein A of Borrelia burgdorferi. Infection and Immunity 63(4): 1356-1361

The gene for the major outer surface protein A (OspA) from several clinically obtained strains of Borrelia burgdorferi, the cause of Lyme disease, has been cloned, sequenced, and expressed in Escherichia coli by using a T7-based expression system (J. J. Dunn, B. N. Lade, and A. G. Barbour, Protein Expr. Purif. 1:159-168, 1990). All of the OspAs have a single conserved tryptophan at residue 216 or, in some cases, 217; however, the region of the protein flanking the tryptophan is hypervariable, as determined by a moving-window population analysis of ospA from 15 European and North American isolates of B. burgdorferi. Epitope-mapping studies using chemically cleaved OspA and a TrpE-OspA fusion have indicated that this hypervariable region is important for immune recognition. Biophysical analysis, including fluorescence and circular dichroism spectroscopy, have indicated that the conserved tryptophan is buried in a hydrophobic environment. Polar amino acid side chains flanking the tryptophan are likely to be exposed to the hydrophilic solvent. The hypervariability of these solvent-exposed amino acid residues may contribute to the antigenic variation in OspA. To test this, we have performed site-directed mutagenesis to replace some of the potentially exposed amino acid side chains in the B31 protein with the analogous residues of a Borrelia garinii strain, K48. The altered proteins were then analyzed by Western blot (immunoblot) with monoclonal antibodies which bind OspA on the surface of the intact B31 spirochete. Our results indicate that specific amino acid changes near the tryptophan can abolish the reactivity of OspA to these monoclonal antibodies, which is an important consideration in the design of vaccines based on recombinant OspA.

(PDF 0-2 workdays service)

Accession: 002863309

PMID: 7890394

Submit PDF Full Text: Here


Submit PDF Full Text

No spam - Every submission is manually reviewed

Due to poor quality, we do not accept files from Researchgate

Submitted PDF Full Texts will always be free for everyone
(We only charge for PDFs that we need to acquire)

Select a PDF file:
Close
Close

Related references

Pal, S.; Luke, C.J.; Barbour, A.G.; Peterson, E.M.; de la Maza, L.M., 2003: Immunization with the Chlamydia trachomatis major outer membrane protein, using the outer surface protein A of Borrelia burgdorferi as an adjuvant, can induce protection against a chlamydial genital challenge. Two strains of mice C3H/HeN (H-2(k)) and BALB/c (H-2(d)) were immunized with the Chlamydia trachomatis mouse pneumonitis (MoPn) major outer membrane protein (MOMP) using the Borrelia burgdorferi outer surface protein A (OspA) as an adjuvant. As a...

Becker, M.; Bunikis, J.; Lade, B.; Dunn, J.J.; Barbour, A.G.; Lawson, C.L., 2001: X-ray crystal structures of the C-terminal domain of Outer Surface Protein B of Borrelia burgdorferi alone and in complex with a bactericidal FAB. Biophysical Journal 80(1 Part 2): 60a, January

Wilske, B.; Preac-Mursic, V.; Jauris, S.; Hofmann, A.; Pradel, I.; Soutschek, E.; Schwab, E.; Will, G.; Wanner, G., 1993: Immunological and molecular polymorphisms of OspC, an immunodominant major outer surface protein of Borrelia burgdorferi. The gene of the immunodominant major protein pC of Borrelia burgdorferi was previously cloned and sequenced (R. Fuchs, S. Jauris, F. Lottspeich, V. Preac-Mursic, B. Wilske, and E. Soutschek, Mol. Microbiol. 6:503-509, 1992). pC is abundantly expre...

Abolhassani, M., 2000: Identification of two epitopes on the outer surface protein A of the Lyme disease spirochete Borrelia burgdorferi. A murine IgM monoclonal antibody (MA-2C6) with kappa -light chains directed against an antigenic determinant of outer surface protein A (OspA) of the Lyme disease spirochaete, Borrelia burgdorferi, is produced. This antibody could bind specificall...

Rosa, P.A.; Hogan, D.; Margolis, N., 1992: Molecular analysis of the major outer surface protein locus from a divergent Borrelia burgdorferi isolate from Europe. Schutzer, S E [Editor] Current Communications in Cell and Molecular Biology; Lyme disease: Molecular and immunologic approaches : 95-110

Schubach, W.H.; Mudri, S.; Dattwyler, R.J.; Luft, B.J., 1991: Mapping antibody-binding domains of the major outer surface membrane protein (OspA) of Borrelia burgdorferi. The major outer surface membrane protein of Borrelia burgdorferi, OspA, is one of several antigens recognized by sera from some patients in the chronic phase of Lyme borreliosis. We have expressed the OspA open reading frame in Escherichia coli an...

Kee, S.; Hwang, K.J.; Oh, H.B.; Park, K.S., 1994: Identification of Borrelia burgdorferi isolated in Korea using outer surface protein A (OspA) serotyping system. Two characteristic strains (935T, 934U) of B. burgdorferi isolated from Ixodes persulcatus and a wild rodent (Apodemus agrarius) in South Korea were selected and analysed by an immunoblot method using the monoclonal antibodies directed to differen...

Mcgrath, B.C.; Dunn, J.J.; France, L.L.; Jaing, W.; Dykhuizen, D.; Polin, D.; Gorgone, G.; Luft, B., 1993: Biochemical and biophysical characterization of the major outer surface protein from North American and European isolates of Borrelia burgdorferi. Ginsberg, H S [Editor], Brown, F [Editor], Chanok, R M [Editor], Lerner, R A [Editor] Vaccines (Cold Spring Harbor); Modern approaches to new vaccines including prevention of AIDS : 365-370

Zhong, W.; Wiesmüller, K.H.; Kramer, M.D.; Wallich, R.; Simon, M.M., 1996: Plasmid DNA and protein vaccination of mice to the outer surface protein A of Borrelia burgdorferi leads to induction of T helper cells with specificity for a major epitope and augmentation of protective IgG antibodies in vivo. Plasmid DNA-based vaccination is an efficient way to evoke various forms of protective immunity in laboratory animals. Our previous experiments have shown that mice immunized with either plasmid DNA encoding the outer surface lipoprotein A (pOspA)...

Pulzova, L.; Flachbartova, Z.; Bencurova, E.; Potocnakova, L.; Comor, L.; Schreterova, E.; Bhide, M., 2016: Identification of B-cell epitopes of Borrelia burgdorferi outer surface protein C by screening a phage-displayed gene fragment library. NlmCategory="UNASSIGNED">Outer surface protein C (OspC) of Borrelia stimulates remarkable immune responses during early infection and is therefore currently considered a leading diagnostic and vaccine candidate. The sensitivity and sp...