Epitope mapping of Burkholderia pseudomallei serine metalloprotease: Identification of serine protease epitope mimics

Shzu Wei Chan, Sheila Nathan

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Filamentous phage random peptide libraries were used to identify the epitopes of Burkholderia pseudomallei protease by panning against IgG polyclonal sera that exhibited protease neutralizing properties. The isolated fusion peptides presented a consensus peptide sequence, TKSMALSG, which closely resembles part of the active site sequence, 435GTSMATPHVAG445, of B. pseudomallei serine metalloprotease. By comparing the consensus sequence, TKSMALSG, with the predicted three-dimensional molecular model of B. pseudomallei serine metalloprotease, it appears that the potential antibody binding epitope was buried within the molecule. This active site was conformational whereby one continuous sub-region (SMA) was located between two discontinuous sub-regions, supplied by the flanking residues in the same polypeptide. All phages selected from the biopanning with IgG polyclonal sera showed good binding towards the polyclonal antibodies when compared to the negative control. In addition, these peptide-bearing phages showed competitive inhibition of B. pseudomallei serine metalloprotease binding to the polyclonal IgG.

Original languageEnglish
Pages (from-to)37-44
Number of pages8
JournalFEMS Immunology and Medical Microbiology
Volume43
Issue number1
DOIs
Publication statusPublished - 1 Jan 2005

Fingerprint

Burkholderia pseudomallei
Epitope Mapping
Metalloproteases
Serine Proteases
Serine
Epitopes
Bacteriophages
Peptides
Immunoglobulin G
Consensus Sequence
Catalytic Domain
Peptide Hydrolases
Peptide Library
Molecular Models
Antibodies
Serum

Keywords

  • Burkholderia pseudomallei
  • Epitope mapping
  • Molecular modeling
  • Phage display

ASJC Scopus subject areas

  • Immunology
  • Microbiology
  • Infectious Diseases

Cite this

@article{ea2016d318ba4541b9851a4edd852bcd,
title = "Epitope mapping of Burkholderia pseudomallei serine metalloprotease: Identification of serine protease epitope mimics",
abstract = "Filamentous phage random peptide libraries were used to identify the epitopes of Burkholderia pseudomallei protease by panning against IgG polyclonal sera that exhibited protease neutralizing properties. The isolated fusion peptides presented a consensus peptide sequence, TKSMALSG, which closely resembles part of the active site sequence, 435GTSMATPHVAG445, of B. pseudomallei serine metalloprotease. By comparing the consensus sequence, TKSMALSG, with the predicted three-dimensional molecular model of B. pseudomallei serine metalloprotease, it appears that the potential antibody binding epitope was buried within the molecule. This active site was conformational whereby one continuous sub-region (SMA) was located between two discontinuous sub-regions, supplied by the flanking residues in the same polypeptide. All phages selected from the biopanning with IgG polyclonal sera showed good binding towards the polyclonal antibodies when compared to the negative control. In addition, these peptide-bearing phages showed competitive inhibition of B. pseudomallei serine metalloprotease binding to the polyclonal IgG.",
keywords = "Burkholderia pseudomallei, Epitope mapping, Molecular modeling, Phage display",
author = "Chan, {Shzu Wei} and Sheila Nathan",
year = "2005",
month = "1",
day = "1",
doi = "10.1016/j.femsim.2004.06.025",
language = "English",
volume = "43",
pages = "37--44",
journal = "Pathogens and Disease",
issn = "2049-632X",
publisher = "John Wiley & Sons Inc.",
number = "1",

}

TY - JOUR

T1 - Epitope mapping of Burkholderia pseudomallei serine metalloprotease

T2 - Identification of serine protease epitope mimics

AU - Chan, Shzu Wei

AU - Nathan, Sheila

PY - 2005/1/1

Y1 - 2005/1/1

N2 - Filamentous phage random peptide libraries were used to identify the epitopes of Burkholderia pseudomallei protease by panning against IgG polyclonal sera that exhibited protease neutralizing properties. The isolated fusion peptides presented a consensus peptide sequence, TKSMALSG, which closely resembles part of the active site sequence, 435GTSMATPHVAG445, of B. pseudomallei serine metalloprotease. By comparing the consensus sequence, TKSMALSG, with the predicted three-dimensional molecular model of B. pseudomallei serine metalloprotease, it appears that the potential antibody binding epitope was buried within the molecule. This active site was conformational whereby one continuous sub-region (SMA) was located between two discontinuous sub-regions, supplied by the flanking residues in the same polypeptide. All phages selected from the biopanning with IgG polyclonal sera showed good binding towards the polyclonal antibodies when compared to the negative control. In addition, these peptide-bearing phages showed competitive inhibition of B. pseudomallei serine metalloprotease binding to the polyclonal IgG.

AB - Filamentous phage random peptide libraries were used to identify the epitopes of Burkholderia pseudomallei protease by panning against IgG polyclonal sera that exhibited protease neutralizing properties. The isolated fusion peptides presented a consensus peptide sequence, TKSMALSG, which closely resembles part of the active site sequence, 435GTSMATPHVAG445, of B. pseudomallei serine metalloprotease. By comparing the consensus sequence, TKSMALSG, with the predicted three-dimensional molecular model of B. pseudomallei serine metalloprotease, it appears that the potential antibody binding epitope was buried within the molecule. This active site was conformational whereby one continuous sub-region (SMA) was located between two discontinuous sub-regions, supplied by the flanking residues in the same polypeptide. All phages selected from the biopanning with IgG polyclonal sera showed good binding towards the polyclonal antibodies when compared to the negative control. In addition, these peptide-bearing phages showed competitive inhibition of B. pseudomallei serine metalloprotease binding to the polyclonal IgG.

KW - Burkholderia pseudomallei

KW - Epitope mapping

KW - Molecular modeling

KW - Phage display

UR - http://www.scopus.com/inward/record.url?scp=11144276434&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=11144276434&partnerID=8YFLogxK

U2 - 10.1016/j.femsim.2004.06.025

DO - 10.1016/j.femsim.2004.06.025

M3 - Article

C2 - 15607634

AN - SCOPUS:11144276434

VL - 43

SP - 37

EP - 44

JO - Pathogens and Disease

JF - Pathogens and Disease

SN - 2049-632X

IS - 1

ER -