Isolation and characterization of human cervical-mucus glycoproteins.
Mucus glycoproteins (mucins) were extracted from human cervical pregnancy mucus by 6 M-guanidinium chloride in the presence of proteinase inhibitors. Purification was subsequently achieved by isopycnic density-gradient centrifugation in CsCl/ guanidinium chloride gradients. The purified macromolecules represented approx. 85% of the total and were devoid of nucleic acids and proteins, as judged by analytical density-gradient centrifugation, disc electrophoresis and u.v. spectroscopy. Sedimentation-velocity centrifugation revealed a single unimodal peak with S20,W 50.1S in 0.2M-NaCl and 37.0S in 6 M-guanidinium chloride. Molecular weights obtained by light-scattering were 9.7 X 10(6) and 5.9 X 10(6) in 0.2M-NaCl and 6 M-guanidinium chloride respectively. The chemical analyses were typical of those of epithelial mucins. The macromolecules contained approx. 20% (w/w) of protein, and 65% (w/w) was accounted for as carbohydrate. Serine and threonine constituted 32 mol/100 mol and proline 10 mol/100 mol of the amino acids. The major sugars found were N-acetylglucosamine (12.8%), N-acetylgalactosamine (9.7%), galactose (18.7%), sialic acid (15.0%) and fucose (7.5%).
D000596:Amino Acids / Q000032:analysis; D002241:Carbohydrates / Q000032:analysis; D002499:Centrifugation, Density Gradient; D002582:Cervix Mucus / Q000032:analysis*; D055598:Chemical Phenomena; D002621:Chemistry; D002850:Chromatography, Gel; D005260:Female; D006801:Humans; D008970:Molecular Weight; D009077:Mucins / Q000302:isolation & purification; D011247:Pregnancy; D012542:Scattering, Radiation; D012694:Serine / Q000032:analysis; D013912:Threonine / Q000032:analysis
D000596:Amino Acids; D002241:Carbohydrates; D009077:Mucins; D013912:Threonine; D012694:Serine