Production of the entire extracellular domain of BP180 (type XVII collagen) by baculovirus expression

Yasuki Hata, Yoshiko Fujii, Kazuyuki Tsunoda, Masayuki Amagai

研究成果: Article

21 引用 (Scopus)

抄録

Bullous pemphigoid (BP) is an acquired autoimmune skin disease, and its target antigens are a 230 kDa plaque protein (BP230) and a 180 kDa transmembrane protein with interrupted collagenous domains (BP180, type XVII collagen), which localize at the hemidesmosome. In this study we have attempted to express the entire extracellular domain of BP180 (rBP180EC) as a secreted protein by baculovirus expression. Seventy out of 83 BP sera (84.4%) showed positive reactivity against rBP180EC by immunoblot analysis, and 56 out of 83 BP sera (67.5%) were positive against rBP180EC by ELISA. These figures were comparable with those when a bacterial recombinant protein encoding the NC16a domain of BP180 (rNC16a) was used as an antigen source. Reactivity of BP sera against rBP180EC by ELISA was completely abolished or significantly reduced by immunocompetition with rNC16a in 11 out of 14 BP sera tested, while the reactivity was not altered in the rest of the three sera. These findings indicate that the NC16a domain represents the major epitopes on the extracellular domain of BP180, although there are some other minor epitopes outside of NC16a which are uniquely expressed by rBP180EC. rBP180EC will be useful to develop a diagnostic tool for BP as well as to dissect a molecular role for BP180 in interactions of keratinocytes with epidermal basement membrane. (C) 2000 Elsevier Science Ireland Ltd.

元の言語English
ページ(範囲)183-190
ページ数8
ジャーナルJournal of Dermatological Science
23
発行部数3
DOI
出版物ステータスPublished - 2000 8

Fingerprint

Bullous Pemphigoid
Baculoviridae
Epitopes
Antigens
Serum
Proteins
Recombinant Proteins
Skin
Enzyme-Linked Immunosorbent Assay
Hemidesmosomes
Bacterial Proteins
Keratinocytes
Skin Diseases
Basement Membrane
Autoimmune Diseases
collagen type XVII

ASJC Scopus subject areas

  • Dermatology

これを引用

@article{46cc37bda8c4460587f3d95de69125a3,
title = "Production of the entire extracellular domain of BP180 (type XVII collagen) by baculovirus expression",
abstract = "Bullous pemphigoid (BP) is an acquired autoimmune skin disease, and its target antigens are a 230 kDa plaque protein (BP230) and a 180 kDa transmembrane protein with interrupted collagenous domains (BP180, type XVII collagen), which localize at the hemidesmosome. In this study we have attempted to express the entire extracellular domain of BP180 (rBP180EC) as a secreted protein by baculovirus expression. Seventy out of 83 BP sera (84.4{\%}) showed positive reactivity against rBP180EC by immunoblot analysis, and 56 out of 83 BP sera (67.5{\%}) were positive against rBP180EC by ELISA. These figures were comparable with those when a bacterial recombinant protein encoding the NC16a domain of BP180 (rNC16a) was used as an antigen source. Reactivity of BP sera against rBP180EC by ELISA was completely abolished or significantly reduced by immunocompetition with rNC16a in 11 out of 14 BP sera tested, while the reactivity was not altered in the rest of the three sera. These findings indicate that the NC16a domain represents the major epitopes on the extracellular domain of BP180, although there are some other minor epitopes outside of NC16a which are uniquely expressed by rBP180EC. rBP180EC will be useful to develop a diagnostic tool for BP as well as to dissect a molecular role for BP180 in interactions of keratinocytes with epidermal basement membrane. (C) 2000 Elsevier Science Ireland Ltd.",
keywords = "Baculovirus expression, BP180 (type XVII collagen), Entire extracellular domain, Production",
author = "Yasuki Hata and Yoshiko Fujii and Kazuyuki Tsunoda and Masayuki Amagai",
year = "2000",
month = "8",
doi = "10.1016/S0923-1811(00)00074-8",
language = "English",
volume = "23",
pages = "183--190",
journal = "Journal of Dermatological Science",
issn = "0923-1811",
publisher = "Elsevier Ireland Ltd",
number = "3",

}

TY - JOUR

T1 - Production of the entire extracellular domain of BP180 (type XVII collagen) by baculovirus expression

AU - Hata, Yasuki

AU - Fujii, Yoshiko

AU - Tsunoda, Kazuyuki

AU - Amagai, Masayuki

PY - 2000/8

Y1 - 2000/8

N2 - Bullous pemphigoid (BP) is an acquired autoimmune skin disease, and its target antigens are a 230 kDa plaque protein (BP230) and a 180 kDa transmembrane protein with interrupted collagenous domains (BP180, type XVII collagen), which localize at the hemidesmosome. In this study we have attempted to express the entire extracellular domain of BP180 (rBP180EC) as a secreted protein by baculovirus expression. Seventy out of 83 BP sera (84.4%) showed positive reactivity against rBP180EC by immunoblot analysis, and 56 out of 83 BP sera (67.5%) were positive against rBP180EC by ELISA. These figures were comparable with those when a bacterial recombinant protein encoding the NC16a domain of BP180 (rNC16a) was used as an antigen source. Reactivity of BP sera against rBP180EC by ELISA was completely abolished or significantly reduced by immunocompetition with rNC16a in 11 out of 14 BP sera tested, while the reactivity was not altered in the rest of the three sera. These findings indicate that the NC16a domain represents the major epitopes on the extracellular domain of BP180, although there are some other minor epitopes outside of NC16a which are uniquely expressed by rBP180EC. rBP180EC will be useful to develop a diagnostic tool for BP as well as to dissect a molecular role for BP180 in interactions of keratinocytes with epidermal basement membrane. (C) 2000 Elsevier Science Ireland Ltd.

AB - Bullous pemphigoid (BP) is an acquired autoimmune skin disease, and its target antigens are a 230 kDa plaque protein (BP230) and a 180 kDa transmembrane protein with interrupted collagenous domains (BP180, type XVII collagen), which localize at the hemidesmosome. In this study we have attempted to express the entire extracellular domain of BP180 (rBP180EC) as a secreted protein by baculovirus expression. Seventy out of 83 BP sera (84.4%) showed positive reactivity against rBP180EC by immunoblot analysis, and 56 out of 83 BP sera (67.5%) were positive against rBP180EC by ELISA. These figures were comparable with those when a bacterial recombinant protein encoding the NC16a domain of BP180 (rNC16a) was used as an antigen source. Reactivity of BP sera against rBP180EC by ELISA was completely abolished or significantly reduced by immunocompetition with rNC16a in 11 out of 14 BP sera tested, while the reactivity was not altered in the rest of the three sera. These findings indicate that the NC16a domain represents the major epitopes on the extracellular domain of BP180, although there are some other minor epitopes outside of NC16a which are uniquely expressed by rBP180EC. rBP180EC will be useful to develop a diagnostic tool for BP as well as to dissect a molecular role for BP180 in interactions of keratinocytes with epidermal basement membrane. (C) 2000 Elsevier Science Ireland Ltd.

KW - Baculovirus expression

KW - BP180 (type XVII collagen)

KW - Entire extracellular domain

KW - Production

UR - http://www.scopus.com/inward/record.url?scp=0033847681&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033847681&partnerID=8YFLogxK

U2 - 10.1016/S0923-1811(00)00074-8

DO - 10.1016/S0923-1811(00)00074-8

M3 - Article

C2 - 10959044

AN - SCOPUS:0033847681

VL - 23

SP - 183

EP - 190

JO - Journal of Dermatological Science

JF - Journal of Dermatological Science

SN - 0923-1811

IS - 3

ER -