Characterization of 230-kD bullous pemphigoid antigen associated with the detergent-insoluble fraction of cultured keratinocytes.
The 230-kD protein identified by antibodies from patients with bullous pemphigoid (BP) has a dual location in cultured normal human epidermal keratinocytes: part in a high-speed supernatant of homogenized cells and part in a particulate fraction, where it is resistant to extraction by non-ionic detergent or mild base. Antibodies were affinity purified from the particulate 230-kD BP antigen, which can be extracted in the presence of urea. The affinity-purified antibodies bind not only the cytosolic 230-kD protein, showing that it is related, if not identical, to the particulate form, but also produce a discontinuous granular pattern by indirect immunofluorescence in the basement membrane zone of rabbit esophagus. In stratifying epidermal cultures, expression of the 230-kD BP antigen is limited to basal cells. These data are consistent with 230-kD BP antigen involvement in keratinocyte basal cell interaction with extracellular matrix and indicate that the cultured cell may provide a useful model for analysis of 230-kD BP antigen function.
Thacher
S M
SM
Department of Medical Biochemistry and Genetics, Texas A & M College of Medicine, College Station 77843.
Hefti
P L
PL
D000818:Animals; D001324:Autoantigens / Q000302:isolation & purification*; D001485:Basement Membrane / Q000166:cytology; D002352:Carrier Proteins; D002478:Cells, Cultured; D002850:Chromatography, Gel; D003094:Collagen; D003598:Cytoskeletal Proteins; D003902:Detergents; D000071821:Dystonin; D004947:Esophagus / Q000166:cytology; D005455:Fluorescent Antibody Technique; D006801:Humans; D015151:Immunoblotting; D007231:Infant, Newborn; D015603:Keratinocytes / Q000737:chemistry* / Q000166:cytology; D008297:Male; D008970:Molecular Weight; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D010391:Pemphigoid, Bullous / Q000276:immunology; D011817:Rabbits; D000097805:Collagen Type XVII
D001324:Autoantigens; D002352:Carrier Proteins; D003094:Collagen; D003598:Cytoskeletal Proteins; D003902:Detergents; D000071821:Dystonin; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D000097805:Collagen Type XVII; C486268:DST protein, human