Direct interaction between the intracellular domains of bullous pemphigoid antigen 2 (BP180) and beta 4 integrin, hemidesmosomal components of basal keratinocytes.
Bullous pemphigoid antigen 2 (BPAG2/BP180, also known as type XVII collagen) and alpha 6 beta 4 integrin are both transmembrane proteins and hemidesmosomal components of basal keratinocytes. In this study, using yeast two-hybrid system, we demonstrate direct protein-protein interaction between the intracellular domains of BP180 and the beta 4 integrin subunit. Detailed analysis revealed that a bait construct spanning amino acids 13-89 of BP180 contained sufficient information for the protein protein interaction, but further deletion of 13 amino-terminal amino acids, which eliminates a predicted beta-sheet, abolished the interaction. The intracellular domain of the beta 4 integrin subunit contains two pairs of fibronectin type III (FNIII) repeats separated by a connecting segment. Series of expression constructs, sequentially deleting each domain, revealed that the connecting segment, the second pair of FNIII repeats and the tail region of the beta 4 integrin subunit were necessary for the interaction with BP180 in yeast two-hybrid system.
Aho
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Department of Dermatology and Cutaneous Biology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Uitto
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J
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000954:Antigens, Surface / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001324:Autoantigens / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003598:Cytoskeletal Proteins; D004268:DNA-Binding Proteins; D000071821:Dystonin; D005353:Fibronectins / Q000737:chemistry; D006801:Humans; D039161:Integrin alpha6beta4; D016023:Integrins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015603:Keratinocytes / Q000737:chemistry* / Q000648:ultrastructure; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D016133:Polymerase Chain Reaction; D017433:Protein Structure, Secondary; D012194:RNA-Directed DNA Polymerase; D011993:Recombinant Fusion Proteins; D012091:Repetitive Sequences, Nucleic Acid; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000235:genetics; D000097805:Collagen Type XVII
D000954:Antigens, Surface; D001324:Autoantigens; D002352:Carrier Proteins; D003094:Collagen; D003598:Cytoskeletal Proteins; D004268:DNA-Binding Proteins; D000071821:Dystonin; D005353:Fibronectins; D039161:Integrin alpha6beta4; D016023:Integrins; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D012194:RNA-Directed DNA Polymerase; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D000097805:Collagen Type XVII; C486268:DST protein, human; C052130:GAL4 protein, S cerevisiae