Molecular heterogeneity of human motilinlike immunoreactivity explained by the processing of prepromotilin.
Results of studies on the molecular forms of canine motilin suggest that this hormone might be synthesized as a higher molecular weight precursor, as is the case for most other biologically active peptides. Chromatographic analysis of human duodenal mucosa extracts and of human serum, using motilin-specific antibodies, also shows the presence of multiple forms of motilinlike immunoreactivity. Cell free translation of human duodenal messenger ribonucleic acid (mRNA) and immunoprecipitation of nascent peptides with motilin antibodies confirm that motilin is synthesized as a 14-15-kilodalton polypeptide precursor. Sequence of the human intestinal motilin complementary deoxyribonucleic acid (cDNA) demonstrates that this precursor bears a 25-amino acid signal peptide, a single dibasic cleavage site immediately following the 22 amino acids of human motilin, and a 65-amino acid polypeptide (motilin-related peptide) following this dibasic processing site. Southern analysis of human genomic kilobases DNA demonstrates that only one motilin gene is contained within 3.5-4 of human genomic DNA. Northern analysis of human intestinal RNA reveals one species of motilin mRNA of an estimated 700 nucleotides in length. These results suggest that (a) only one gene encodes the synthesis of human motilin in the different tissues where this polypeptide is synthesized; (b) human intestinal motilin is translated from one mRNA species and the resulting precursor is processed within the duodenal mucosa by sequential proteolytic cleavage first at a site within the motilin-related peptide, which results in the liberation of a 6-kilodalton form of motilinlike immunoreactivity, and then at the dibasic amino acid cleavage site, thus freeing motilin 22 from its precursor.
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Centre de Recherche Clinique André-Viallet, Hôpital Saint-Luc, Montreal, Québec, Canada.
Boileau
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Poitras
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P
Lahaie
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RG
D000595:Amino Acid Sequence; D004247:DNA / Q000235:genetics; D004386:Duodenum / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007413:Intestinal Mucosa / Q000378:metabolism; D008969:Molecular Sequence Data; D009037:Motilin / Q000235:genetics*; D011498:Protein Precursors / Q000235:genetics*; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic
D011498:Protein Precursors; D012333:RNA, Messenger; C058325:prepromotilin; D009037:Motilin; D004247:DNA