The extracellular domain of BPAG2 localizes to anchoring filaments and its carboxyl terminus extends to the lamina densa of normal human epidermal basement membrane

Takuji Masunaga, Hiroshi Shimizu, Carole Yee, Luca Borradori, Zelmira Lazarova, Takeji Nishikawa, Kim B. Yancey

Research output: Contribution to journalArticle

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Abstract

Bullous pemphigoid antigen 2 (BPAG2) is a 180 kDa type II transmembrane protein associated with hemidesmosomes (HDs) in basal keratinocytes. To better understand how BPAG2 promotes keratinocyte adhesion to epidermal basement membrane (BM), purified IgG against a baculovirus-encoded recombinant was used to localize its carboxyl terminus in human skin by immunogold electron microscopy (IEM). A 2.1-kb BPAG2 cDNA encoding the distal extracellular domain and carboxyl terminus of BPAG2 was used in a baculovirus expression system to create virus that produced a 70-kDa recombinant form of BPAG2 (BV4). BV4 was purified, characterized, and used to raise high-titer specific rabbit IgG. Purified anti-BV4 IgG bound the epidermal side of 1 M NaC1 split skin and bound only BPAG2 on immunoblots containing extracts of human keratinocytes. In IEM studies of pre- and post-embedded skin, the distal ectodomain of BPAG2 localized beneath HDs in basal keratinocytes; there was no evidence of BPAG2 beneath melanocytes. Anti-BV4 IgG extensively bound anchoring filaments on the epidermal side of 1 M NaC1 split skin; this staining extended along anchoring filaments to their ends. In post-embedded skin, the carboxyl terminus of BPAG2 was localized within the lamina densa, 41 nm (mean of 400 determinations) beneath plasma membranes of basal keratinocytes. BPAG2 thus extends from the intracellular HD plaque of basal keratinocytes to the lamina densa of human epidermal BM.

Original languageEnglish (US)
Pages (from-to)200-206
Number of pages7
JournalJournal of Investigative Dermatology
Volume109
Issue number2
StatePublished - 1997

Fingerprint

Bullous Pemphigoid
Basement Membrane
Antigens
Keratinocytes
Hemidesmosomes
Skin
Baculoviridae
Electron microscopy
Electron Microscopy
Immunoglobulin G
Melanocytes
Cell membranes
Viruses
Adhesion
Complementary DNA
Cell Membrane
Staining and Labeling
Rabbits

Keywords

  • Collagen
  • Hemidesmosomes
  • Laminin 5

ASJC Scopus subject areas

  • Dermatology

Cite this

The extracellular domain of BPAG2 localizes to anchoring filaments and its carboxyl terminus extends to the lamina densa of normal human epidermal basement membrane. / Masunaga, Takuji; Shimizu, Hiroshi; Yee, Carole; Borradori, Luca; Lazarova, Zelmira; Nishikawa, Takeji; Yancey, Kim B.

In: Journal of Investigative Dermatology, Vol. 109, No. 2, 1997, p. 200-206.

Research output: Contribution to journalArticle

Masunaga, Takuji ; Shimizu, Hiroshi ; Yee, Carole ; Borradori, Luca ; Lazarova, Zelmira ; Nishikawa, Takeji ; Yancey, Kim B. / The extracellular domain of BPAG2 localizes to anchoring filaments and its carboxyl terminus extends to the lamina densa of normal human epidermal basement membrane. In: Journal of Investigative Dermatology. 1997 ; Vol. 109, No. 2. pp. 200-206.
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