PMID: 10022517Feb 18, 1999

180-kD bullous pemphigoid antigen/type XVII collagen: tissue-specific expression and molecular interactions with keratin 18

Journal of Cellular Biochemistry
S Aho, Jouni Uitto

Abstract

The 180-kD bullous pemphigoid antigen (BPAG2) is a hemidesmosomal transmembrane protein, also known as type XVII collagen. In this study, potential interactions of BPAG2 with other proteins expressed in epidermal keratinocytes were explored by yeast two-hybrid system using the amino-terminal intracellular domain of BPAG2 as a bait. Several independent interacting clones encoding keratin 18 (K18) were identified when the keratinocyte cDNA library, cloned into the yeast two-hybrid activation domain vector, was screened. The peptide sequence responsible for the interaction of BPAG2 was restricted to amino acids 15-25, and substitution of a valine residue in the middle of this sequence by a proline (V23P) by site-directed mutagenesis abolished the interaction. Further examination of the K18 sequences by restricted cDNA constructs in yeast two-hybrid system identified a carboxyl-terminal segment corresponding to helix 2B domain as critical for BPAG2 binding. The interaction of BPAG2/K18 was confirmed by an in vitro protein-protein interaction assay, which also confirmed that normal human keratinocytes express K18 in culture. The tissue specific expression of BPAG2 was first examined using a multi-tissue RNA blot. Human multiple tiss...Continue Reading

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Related Concepts

Fetal Kidney
Ovarian Diseases
Tumor Cells, Uncertain Whether Benign or Malignant
Yeast Two-Hybrid System Techniques
Benign Neoplasm of Testis
Fetal Structures
Molecular Helix
Chimeric Proteins, Recombinant
Epithelial Ovarian Cancer
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