Evidence for an α-helical epitope on outer surface protein A from the Lyme disease spirochete, Borrelia burgdorferi

An application of steady-state and time-resolved fluorescence quenching techniques

Louisa L. France, Jan Kieleczawa, John J. Dunn, Benjamin J. Luft, Geoffrey Hind, John C. Sutherland

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Outer surface protein A (OspA) is a major antigen of Borrelia burgdorferi, the etiological agent of Lyme disease. A recombinant form of OspA (OspA-257) from B. burgdorferi, strain B31, contains 257 amino acids and a single tryptophan residue at position 216 (Trp-216). Mapping studies indicate that Trp-216 is involved in the epitope for the agglutinating monoclonal antibody 105.5 However, the fluorescence emission maximum of the native protein is 330 nm, indicating that Trp-216 is not solvent-exposed. Primary structure analysis suggests and α-helical conformation for residues approx. 204-217, which, if located on the protein surface, would allow Trp-216 to be buried, while leaving hydrophilic residues on the opposite side of the helix exposed. This helix would plate Lys-212 within approx. 6 Å of Trp-216; the presence of such a positively-charged residue can, in principle, be ascertained from fluorescence quenching studies. Stern-Volmer plots confirm that Trp-216 is indeed buried in the native protein, but is readily accessible to the small polar quencher, Cs+, while the minor component undergoes static quenching by I-, indicating the proximity of a positively-charged residue. These data are consistent with the existence of an α-helix from residues 204-217 in the predicted orientation at the protein surface, hence indicating the structure of the antigentic determinant.

Original languageEnglish (US)
Pages (from-to)287-296
Number of pages10
JournalBiochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
Volume1202
Issue number2
DOIs
StatePublished - Oct 6 1993
Externally publishedYes

Fingerprint

Borrelia burgdorferi
Tryptophan
Epitopes
Quenching
Fluorescence
Membrane Proteins
Lyme Disease
Conformations
OspA protein
Proteins
Monoclonal Antibodies
Antigens
Amino Acids

Keywords

  • Alpha helix
  • Fluorescence quenching
  • Lyme disease
  • Outer surface protein A
  • Time-resolved fluorescence
  • Tryptophan fluorescence

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Structural Biology

Cite this

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title = "Evidence for an α-helical epitope on outer surface protein A from the Lyme disease spirochete, Borrelia burgdorferi: An application of steady-state and time-resolved fluorescence quenching techniques",
abstract = "Outer surface protein A (OspA) is a major antigen of Borrelia burgdorferi, the etiological agent of Lyme disease. A recombinant form of OspA (OspA-257) from B. burgdorferi, strain B31, contains 257 amino acids and a single tryptophan residue at position 216 (Trp-216). Mapping studies indicate that Trp-216 is involved in the epitope for the agglutinating monoclonal antibody 105.5 However, the fluorescence emission maximum of the native protein is 330 nm, indicating that Trp-216 is not solvent-exposed. Primary structure analysis suggests and α-helical conformation for residues approx. 204-217, which, if located on the protein surface, would allow Trp-216 to be buried, while leaving hydrophilic residues on the opposite side of the helix exposed. This helix would plate Lys-212 within approx. 6 {\AA} of Trp-216; the presence of such a positively-charged residue can, in principle, be ascertained from fluorescence quenching studies. Stern-Volmer plots confirm that Trp-216 is indeed buried in the native protein, but is readily accessible to the small polar quencher, Cs+, while the minor component undergoes static quenching by I-, indicating the proximity of a positively-charged residue. These data are consistent with the existence of an α-helix from residues 204-217 in the predicted orientation at the protein surface, hence indicating the structure of the antigentic determinant.",
keywords = "Alpha helix, Fluorescence quenching, Lyme disease, Outer surface protein A, Time-resolved fluorescence, Tryptophan fluorescence",
author = "France, {Louisa L.} and Jan Kieleczawa and Dunn, {John J.} and Luft, {Benjamin J.} and Geoffrey Hind and Sutherland, {John C.}",
year = "1993",
month = "10",
day = "6",
doi = "10.1016/0167-4838(93)90018-M",
language = "English (US)",
volume = "1202",
pages = "287--296",
journal = "Biochimica et Biophysica Acta - Proteins and Proteomics",
issn = "1570-9639",
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TY - JOUR

T1 - Evidence for an α-helical epitope on outer surface protein A from the Lyme disease spirochete, Borrelia burgdorferi

T2 - An application of steady-state and time-resolved fluorescence quenching techniques

AU - France, Louisa L.

AU - Kieleczawa, Jan

AU - Dunn, John J.

AU - Luft, Benjamin J.

AU - Hind, Geoffrey

AU - Sutherland, John C.

PY - 1993/10/6

Y1 - 1993/10/6

N2 - Outer surface protein A (OspA) is a major antigen of Borrelia burgdorferi, the etiological agent of Lyme disease. A recombinant form of OspA (OspA-257) from B. burgdorferi, strain B31, contains 257 amino acids and a single tryptophan residue at position 216 (Trp-216). Mapping studies indicate that Trp-216 is involved in the epitope for the agglutinating monoclonal antibody 105.5 However, the fluorescence emission maximum of the native protein is 330 nm, indicating that Trp-216 is not solvent-exposed. Primary structure analysis suggests and α-helical conformation for residues approx. 204-217, which, if located on the protein surface, would allow Trp-216 to be buried, while leaving hydrophilic residues on the opposite side of the helix exposed. This helix would plate Lys-212 within approx. 6 Å of Trp-216; the presence of such a positively-charged residue can, in principle, be ascertained from fluorescence quenching studies. Stern-Volmer plots confirm that Trp-216 is indeed buried in the native protein, but is readily accessible to the small polar quencher, Cs+, while the minor component undergoes static quenching by I-, indicating the proximity of a positively-charged residue. These data are consistent with the existence of an α-helix from residues 204-217 in the predicted orientation at the protein surface, hence indicating the structure of the antigentic determinant.

AB - Outer surface protein A (OspA) is a major antigen of Borrelia burgdorferi, the etiological agent of Lyme disease. A recombinant form of OspA (OspA-257) from B. burgdorferi, strain B31, contains 257 amino acids and a single tryptophan residue at position 216 (Trp-216). Mapping studies indicate that Trp-216 is involved in the epitope for the agglutinating monoclonal antibody 105.5 However, the fluorescence emission maximum of the native protein is 330 nm, indicating that Trp-216 is not solvent-exposed. Primary structure analysis suggests and α-helical conformation for residues approx. 204-217, which, if located on the protein surface, would allow Trp-216 to be buried, while leaving hydrophilic residues on the opposite side of the helix exposed. This helix would plate Lys-212 within approx. 6 Å of Trp-216; the presence of such a positively-charged residue can, in principle, be ascertained from fluorescence quenching studies. Stern-Volmer plots confirm that Trp-216 is indeed buried in the native protein, but is readily accessible to the small polar quencher, Cs+, while the minor component undergoes static quenching by I-, indicating the proximity of a positively-charged residue. These data are consistent with the existence of an α-helix from residues 204-217 in the predicted orientation at the protein surface, hence indicating the structure of the antigentic determinant.

KW - Alpha helix

KW - Fluorescence quenching

KW - Lyme disease

KW - Outer surface protein A

KW - Time-resolved fluorescence

KW - Tryptophan fluorescence

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U2 - 10.1016/0167-4838(93)90018-M

DO - 10.1016/0167-4838(93)90018-M

M3 - Article

VL - 1202

SP - 287

EP - 296

JO - Biochimica et Biophysica Acta - Proteins and Proteomics

JF - Biochimica et Biophysica Acta - Proteins and Proteomics

SN - 1570-9639

IS - 2

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