Identification of a second protein product of the gene encoding a human epidermal autoantigen.
A 230 kDa polypeptide component of the hemidesmosome, an epithelial-cell-connective-tissue attachment device, is thought to be involved in cytoskeleton-cell-surface anchorage. This 230 kDa polypeptide is recognized by bullous pemphigoid auto-antibodies and for this reason is generally termed the bullous pemphigoid antigen (BPA). We have identified two distinct mRNA products of the single BPA gene by RACE (rapid amplification of cDNA ends)/PCR techniques. The first of these mRNAs encodes the 230 kDa protein component of the hemidesmosome. A second mRNA lacks over 1800 bases that encode the C-terminus of the 230 kDa protein. We have raised antibodies against a peptide specific to the predicted protein product of this second mRNA. To our surprise this antibody recognizes a protein that migrates at 280 kDa on SDS/PAGE of extracts of a variety of human epidermal cell lines that also express the 230 kDa BPA. Moreover, we have confirmed the co-expression of the 230 and 280 kDa polypeptides in these cells by immunoblotting analyses using a monoclonal antibody preparation directed against a polypeptide encoded by sequence common to both mRNAs transcribed from the BPA gene. Intriguingly, in one non-epidermal tumour line (a pancreatic cell line termed FG), the 280 kDa polypeptide appears to be the only product of the BPA gene. Furthermore, in FG cells the 280 kDa protein is found in association with the intermediate filament cytoskeleton. We discuss our results in relation to control of BPA gene expression and with regard to potential functions of the domains of the protein products of the BPA gene.
Northwestern University Medical School, Chicago, IL 60611.
S B
SB
Hopkinson
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JC
Jones
D017398:Alternative Splicing; D001324:Autoantigens / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D002352:Carrier Proteins; D003094:Collagen; D003598:Cytoskeletal Proteins; D018076:DNA, Complementary / Q000235:genetics; D003896:Desmosomes / Q000737:chemistry*; D000071821:Dystonin; D004817:Epidermis / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D005796:Genes; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D010391:Pemphigoid, Bullous / Q000276:immunology*; D010455:Peptides / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics; D000097805:Collagen Type XVII
D001324:Autoantigens; D002352:Carrier Proteins; D003094:Collagen; D003598:Cytoskeletal Proteins; D018076:DNA, Complementary; D000071821:Dystonin; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D010455:Peptides; D012333:RNA, Messenger; D000097805:Collagen Type XVII; C486268:DST protein, human