Disulfide-bound proteolytic fragments of gastric mucin are 100- and 140-kDa proteins.
Pig gastric mucus was tested for its autodegradative proteolytic degradation at pH 7.0, in the presence or absence of proteinase inhibitors and SDS. Samples of crude mucus were incubated at room temperature for 48 and 96 h in sodium azide stabilized buffer, pH 7. 0, and urea-extracted mucin was purified. Electrophoretically homogenic mucin preparation was reduced and alkylated with iodo[(14)C]acetamide, and analyzed for labeled products. On 7.5% SDS/PAGE protein bands at 80 and 120 kDa were noted, but radioactivity was incorporated into 100- and 140-kDa bands, with increasing intensity from T(0) to T(96), and into high molecular mass mucin subunits. The results confirmed the autodegradative properties of gastric mucin and demonstrated that the 100- and 140-kDa fragments are the main proteolytical products of pig gastric mucin and are disulfide bound with the rest of the molecule.
Minkiewicz-Radziejewska
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I
Department of General and Organic Chemistry, Medical Academy of Bialystok, Bialystok 8, Poland.
Gindzieński
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A
Zwierz
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K
D000818:Animals; D002250:Carbon Radioisotopes; D002850:Chromatography, Gel; D004220:Disulfides; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases; D005752:Gastric Mucins / Q000737:chemistry* / Q000302:isolation & purification; D005753:Gastric Mucosa / Q000737:chemistry; D006031:Glycosylation; D007460:Iodoacetamide; D008958:Models, Molecular; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry*; D011487:Protein Conformation; D013552:Swine
D002250:Carbon Radioisotopes; D004220:Disulfides; D005752:Gastric Mucins; D010446:Peptide Fragments; D010450:Endopeptidases; D007460:Iodoacetamide