Subunit structure of porcine submaxillary mucin.
The structure of a high molecular weight fraction of porcine submaxillary mucin was studied by using degradative techniques. Reduction of disulfide linkages released mucin subunits together with an associated protein(s) of approximately 140 kDa. The molecular weights of the subunits ranged from approximately 0.5 x 10(6) to 2.5 x 10(6). Trypsinization of subunits generated glycosylated domains and small, poorly glycosylated or nonglycosylated tryptic peptides. The glycosylated domains, which have an average molecular weight of approximately 270K, possess an unusual amino acid composition containing only nine different amino acids. The minor amino acids which are absent from the glycosylated domains but which are consistently present in both the mucin and the mucin subunits were recovered in the tryptic peptides. Pronase digestion of the glycosylated domains generated smaller fragments of approximately 17 kDa. Comparing these results to the partial cDNA sequence for porcine submaxillary mucin reported by Timpte et al. [(1988) J. Biol. Chem. 263, 1081-1088] suggests that the glycosylated domains consist of variable numbers of the 81 amino acid tandem repeat observed in the cDNA sequence. Further, the fact that porcine submaxillary mucin contains subunits, link proteins, and glycosylated domains suggests that its structure is similar to that described for cervical and intestinal mucins. Intact mucin, mucin "subunits", and the glycosylated domains are all polydisperse with respect to molecular weight, indicating that mucin polydispersity is due to variability in the number of units linked together as well as to variability in the size of the units.
Department of Pediatrics, Rainbow Babies and Children's Hospital, Case Western Reserve University, Cleveland, Ohio 44106.
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Gupta
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Jentoft
D000818:Animals; D006031:Glycosylation; D008970:Molecular Weight; D009077:Mucins / Q000302:isolation & purification; D010446:Peptide Fragments / Q000302:isolation & purification; D011487:Protein Conformation; D013363:Submandibular Gland / Q000032:analysis; D013552:Swine; D014357:Trypsin
D009077:Mucins; D010446:Peptide Fragments; D014357:Trypsin