title: Studies on the peptide core of human bronchial mucus glycoprotein.
abstract: We have employed thiol-Sepharose chromatography following deglycosylation to analyze the protein core of bronchial epithelial mucus glycoprotein (MGP), isolated by a two stage density gradient ultracentrifugation. Deglycosylation using triflouromethanesulfonic acid resulted in loss of greater than 90% of carbohydrate. The deglycosylated core protein was reduced and the sulfhydryl residues activated with 2-2'dipyridyl disulfide. This preparation was then bound covalently to thiol-Sepharose, and eluted specifically with reducing agents. Our results demonstrate that bronchial MGP contains cysteine residues potentially capable of forming disulfide bonds. Pepsin digestion studies suggest that cysteine residues are present near both the heavily glycosylated region and the naked peptide region. Thiol-Sepharose chromatography resolved several mucin-associated proteins (MAPS) that did not bind to the column. Amino acid analysis showed that the largest of these (200 kDa) is enriched in serine/threonine, like MGP that absorbed to the column: the two smallest (20 kDa and 60 kDa) are similar to the proline rich proteins reported in salivary mucin. These associated proteins, although not linked by disulfide bonds to the MGP, are, nevertheless, tightly bound to it, since they were only recovered after deglycosylation and thiol chromatography.
authors.affiliation: Harvard Medical School, Beth Israel Hospital, Boston, Massachusetts.
authors.forename: C
authors.initials: C
authors.lastname: Gerard
authors.forename: K R
authors.initials: KR
authors.lastname: Bhaskar
authors.forename: N P
authors.initials: NP
authors.lastname: Gerard
authors.forename: J M
authors.initials: JM
authors.lastname: Drazen
authors.forename: L
authors.initials: L
authors.lastname: Reid
mesh_terms: D000818:Animals; D001980:Bronchi; D002846:Chromatography, Affinity; D002847:Chromatography, Agarose; D003545:Cysteine / Q000032:analysis / Q000378:metabolism; D006023:Glycoproteins / Q000032:analysis* / Q000302:isolation & purification; D006031:Glycosylation; D006801:Humans; D010455:Peptides / Q000032:analysis*; D011487:Protein Conformation
chemical_list: D006023:Glycoproteins; D010455:Peptides; C047874:gastric mucus glycoproteins; D003545:Cysteine