Epitope mapping of heat shock protein 60 (GroEL) from Porphyromonas gingivalis

Hiroshi Maeda, Manabu Miyamoto, Susumu Kokeguchi, Takayuki Kono, Fusanori Nishimura, Shogo Takashiba, Yoji Murayama

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Porphyromonas gingivalis, a putative pathogen in human periodontal disease, possesses a 60-kDa heat shock protein (hsp60, GroEL). The GroEL homologs are known to be key molecules in auto-immune reactions because of the sequence similarity with human hsp60. In this study, B-cell epitopes on P. gingivalis GroEL (PgGroEL) were analyzed by both Western immunoblotting with truncated PgGroEL and by the multi-pin synthetic peptide approach. To examine auto-antibody production in periodontitis patients, Western immunoblotting with human gingival fibroblasts was performed. Deletion mutants were constructed from the cloned PgGroEL gene (P. gingivalis groEL), and four C-terminal truncated PgGroEL and one N-terminal truncated PgGroEL were prepared from the deletants. Sera from periodontitis patients reacted with all truncated PgGroEL used in this study. The results suggest that the B-cell epitopes were overlaid throughout PgGroEL. To determine the detailed locations of the B-cell epitope, 84 decapeptides covering the entire PgGroEL were synthesized and the serum IgG response to the peptides was examined. Epitope mapping using the synthetic peptides confirmed that the B-cell epitopes were overlaid throughout the length of PgGroEL and revealed that highly conserved peptides between PgGroEL and human hsp60 were recognized by the serum antibodies. Immuno-reactivity against human gingival fibroblasts was examined with sera from 30 periodontitis patients and 10 periodontally healthy subjects. IgG antibody against the 65-kDa antigen in human gingival fibroblasts (same molecular mass as human hsp60) was detected in two patients. Although IgG production against human hsp60 may be rare case in periodontitis patients, the results of epitope mapping demonstrated the potential of PgGroEL to cause the cross-reactions with human hsp60.

Original languageEnglish
Pages (from-to)219-224
Number of pages6
JournalFEMS Immunology and Medical Microbiology
Volume28
Issue number3
DOIs
Publication statusPublished - 2000

Fingerprint

Chaperonin 60
Epitope Mapping
Porphyromonas gingivalis
B-Lymphocyte Epitopes
Periodontitis
Fibroblasts
Immunoglobulin G
Serum
Western Blotting
Antibodies
Cross Reactions
Periodontal Diseases
Antibody Formation
Healthy Volunteers

Keywords

  • B-cell epitope
  • Heat shock protein 60 (GroEL)
  • Porphyromonas gingivalis

ASJC Scopus subject areas

  • Immunology
  • Microbiology
  • Infectious Diseases

Cite this

Epitope mapping of heat shock protein 60 (GroEL) from Porphyromonas gingivalis. / Maeda, Hiroshi; Miyamoto, Manabu; Kokeguchi, Susumu; Kono, Takayuki; Nishimura, Fusanori; Takashiba, Shogo; Murayama, Yoji.

In: FEMS Immunology and Medical Microbiology, Vol. 28, No. 3, 2000, p. 219-224.

Research output: Contribution to journalArticle

Maeda, Hiroshi ; Miyamoto, Manabu ; Kokeguchi, Susumu ; Kono, Takayuki ; Nishimura, Fusanori ; Takashiba, Shogo ; Murayama, Yoji. / Epitope mapping of heat shock protein 60 (GroEL) from Porphyromonas gingivalis. In: FEMS Immunology and Medical Microbiology. 2000 ; Vol. 28, No. 3. pp. 219-224.
@article{160fbc6e6619455d82df656cb588801c,
title = "Epitope mapping of heat shock protein 60 (GroEL) from Porphyromonas gingivalis",
abstract = "Porphyromonas gingivalis, a putative pathogen in human periodontal disease, possesses a 60-kDa heat shock protein (hsp60, GroEL). The GroEL homologs are known to be key molecules in auto-immune reactions because of the sequence similarity with human hsp60. In this study, B-cell epitopes on P. gingivalis GroEL (PgGroEL) were analyzed by both Western immunoblotting with truncated PgGroEL and by the multi-pin synthetic peptide approach. To examine auto-antibody production in periodontitis patients, Western immunoblotting with human gingival fibroblasts was performed. Deletion mutants were constructed from the cloned PgGroEL gene (P. gingivalis groEL), and four C-terminal truncated PgGroEL and one N-terminal truncated PgGroEL were prepared from the deletants. Sera from periodontitis patients reacted with all truncated PgGroEL used in this study. The results suggest that the B-cell epitopes were overlaid throughout PgGroEL. To determine the detailed locations of the B-cell epitope, 84 decapeptides covering the entire PgGroEL were synthesized and the serum IgG response to the peptides was examined. Epitope mapping using the synthetic peptides confirmed that the B-cell epitopes were overlaid throughout the length of PgGroEL and revealed that highly conserved peptides between PgGroEL and human hsp60 were recognized by the serum antibodies. Immuno-reactivity against human gingival fibroblasts was examined with sera from 30 periodontitis patients and 10 periodontally healthy subjects. IgG antibody against the 65-kDa antigen in human gingival fibroblasts (same molecular mass as human hsp60) was detected in two patients. Although IgG production against human hsp60 may be rare case in periodontitis patients, the results of epitope mapping demonstrated the potential of PgGroEL to cause the cross-reactions with human hsp60.",
keywords = "B-cell epitope, Heat shock protein 60 (GroEL), Porphyromonas gingivalis",
author = "Hiroshi Maeda and Manabu Miyamoto and Susumu Kokeguchi and Takayuki Kono and Fusanori Nishimura and Shogo Takashiba and Yoji Murayama",
year = "2000",
doi = "10.1016/S0928-8244(00)00159-0",
language = "English",
volume = "28",
pages = "219--224",
journal = "Pathogens and Disease",
issn = "2049-632X",
publisher = "John Wiley & Sons Inc.",
number = "3",

}

TY - JOUR

T1 - Epitope mapping of heat shock protein 60 (GroEL) from Porphyromonas gingivalis

AU - Maeda, Hiroshi

AU - Miyamoto, Manabu

AU - Kokeguchi, Susumu

AU - Kono, Takayuki

AU - Nishimura, Fusanori

AU - Takashiba, Shogo

AU - Murayama, Yoji

PY - 2000

Y1 - 2000

N2 - Porphyromonas gingivalis, a putative pathogen in human periodontal disease, possesses a 60-kDa heat shock protein (hsp60, GroEL). The GroEL homologs are known to be key molecules in auto-immune reactions because of the sequence similarity with human hsp60. In this study, B-cell epitopes on P. gingivalis GroEL (PgGroEL) were analyzed by both Western immunoblotting with truncated PgGroEL and by the multi-pin synthetic peptide approach. To examine auto-antibody production in periodontitis patients, Western immunoblotting with human gingival fibroblasts was performed. Deletion mutants were constructed from the cloned PgGroEL gene (P. gingivalis groEL), and four C-terminal truncated PgGroEL and one N-terminal truncated PgGroEL were prepared from the deletants. Sera from periodontitis patients reacted with all truncated PgGroEL used in this study. The results suggest that the B-cell epitopes were overlaid throughout PgGroEL. To determine the detailed locations of the B-cell epitope, 84 decapeptides covering the entire PgGroEL were synthesized and the serum IgG response to the peptides was examined. Epitope mapping using the synthetic peptides confirmed that the B-cell epitopes were overlaid throughout the length of PgGroEL and revealed that highly conserved peptides between PgGroEL and human hsp60 were recognized by the serum antibodies. Immuno-reactivity against human gingival fibroblasts was examined with sera from 30 periodontitis patients and 10 periodontally healthy subjects. IgG antibody against the 65-kDa antigen in human gingival fibroblasts (same molecular mass as human hsp60) was detected in two patients. Although IgG production against human hsp60 may be rare case in periodontitis patients, the results of epitope mapping demonstrated the potential of PgGroEL to cause the cross-reactions with human hsp60.

AB - Porphyromonas gingivalis, a putative pathogen in human periodontal disease, possesses a 60-kDa heat shock protein (hsp60, GroEL). The GroEL homologs are known to be key molecules in auto-immune reactions because of the sequence similarity with human hsp60. In this study, B-cell epitopes on P. gingivalis GroEL (PgGroEL) were analyzed by both Western immunoblotting with truncated PgGroEL and by the multi-pin synthetic peptide approach. To examine auto-antibody production in periodontitis patients, Western immunoblotting with human gingival fibroblasts was performed. Deletion mutants were constructed from the cloned PgGroEL gene (P. gingivalis groEL), and four C-terminal truncated PgGroEL and one N-terminal truncated PgGroEL were prepared from the deletants. Sera from periodontitis patients reacted with all truncated PgGroEL used in this study. The results suggest that the B-cell epitopes were overlaid throughout PgGroEL. To determine the detailed locations of the B-cell epitope, 84 decapeptides covering the entire PgGroEL were synthesized and the serum IgG response to the peptides was examined. Epitope mapping using the synthetic peptides confirmed that the B-cell epitopes were overlaid throughout the length of PgGroEL and revealed that highly conserved peptides between PgGroEL and human hsp60 were recognized by the serum antibodies. Immuno-reactivity against human gingival fibroblasts was examined with sera from 30 periodontitis patients and 10 periodontally healthy subjects. IgG antibody against the 65-kDa antigen in human gingival fibroblasts (same molecular mass as human hsp60) was detected in two patients. Although IgG production against human hsp60 may be rare case in periodontitis patients, the results of epitope mapping demonstrated the potential of PgGroEL to cause the cross-reactions with human hsp60.

KW - B-cell epitope

KW - Heat shock protein 60 (GroEL)

KW - Porphyromonas gingivalis

UR - http://www.scopus.com/inward/record.url?scp=0034426478&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034426478&partnerID=8YFLogxK

U2 - 10.1016/S0928-8244(00)00159-0

DO - 10.1016/S0928-8244(00)00159-0

M3 - Article

C2 - 10865174

AN - SCOPUS:0034426478

VL - 28

SP - 219

EP - 224

JO - Pathogens and Disease

JF - Pathogens and Disease

SN - 2049-632X

IS - 3

ER -