Mouse peripherin isoforms

• Published in: Biology of the cell 2000-09, Vol.92 (6), p.397-407
• Main Authors: Landon, Françoise, Wolff, Annie, de Néchaud, Béatrice
• Format: Article
• Language: English
• Subjects: Animals; Cloning, Molecular; Electrophoresis, Polyacrylamide Gel; Immune Sera - immunology; Immunohistochemistry; intermediate filament; Intermediate Filament Proteins - chemistry; Intermediate Filament Proteins - genetics; Intermediate Filament Proteins - immunology; isoform; Membrane Glycoproteins; Mice; Nerve Tissue - chemistry; Nerve Tissue - metabolism; Nerve Tissue Proteins - chemistry; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - immunology; Neuroblastoma - chemistry; peripherin; Peripherins; Protein Isoforms - chemistry; Protein Isoforms - genetics; Protein Isoforms - immunology; Recombinant Proteins - biosynthesis; Recombinant Proteins - isolation & purification; RNA, Messenger - chemistry; RNA, Messenger - metabolism; Tissue Distribution; Tumor Cells, Cultured - chemistry
• ISSN: 0248-4900; 1768-322X
• DOI: 10.1016/S0248-4900(00)01099-6

Three distinct mRNAs have been shown to be produced by alternative splicing from the unique mouse peripherin gene. They generate three translation products, one major form, Pe–58, and two minor forms, Pe–56 which possess a shorter C-terminal sequence, and Pe–61 in which an additional sequence has been inserted in the central rod domain (Landon et al., 1989, EMBO J. 8, 1719–1726). In this study, the simultaneous occurrence of multiple transcripts in murine nervous tissues and neuroblastoma cell lines was shown by PCR amplification of fragments overlapping the sites of alternative splicing. Recombinant peripherin isoforms were purified from E. coli expressing full-length cDNAs. Rabbit antisera were raised against synthetic peptides mimicking parts of the two C-terminal sequences and of the inserted sequence of Pe–61 and were immunoadsorbed until they became monoreactive. By western blot analysis, the peripherin isoforms were localised in neuroblastoma NB2a cell lysates and detergent insoluble fractions separated by two-dimensional electrophoresis. In addition, each isoform was resolved into several charge variants. At the cellular level, each antibody decorated the filament array of the NB2a cells, suggesting the participation of the minor peripherin isoforms in the intermediate filament network.