Characterization of human middle ear mucus glycoprotein in chronic secretory otitis media (CSOM).
Middle ear effusion was obtained from children with chronic secretory otitis media undergoing myringotomy. The effusions contained about 120 mg/ml non-dialysable solids, of which 18-31% was mucus glycoprotein. The purified mucus glycoprotein had a composition characteristic of other mucus glycoproteins. Amino acid analysis of the glycoprotein indicates a protein core consisting of glycosylated regions resistant to proteolysis and non-glycosylated regions susceptible to proteolysis. Analysis of the mucus glycoprotein by gel filtration on Sepharose 2B showed that reduction caused a decrease in hydrodynamic size and proteolysis caused a further decrease. The difference was confirmed by sedimentation coefficient and viscosity measurements. The reduced glycoprotein had an intrinsic viscosity of 0.113 ml/mg and an S0(20) of 15.2S compared to a value of 0.018 ml/mg and 9.6S for the proteolytically digested glycoprotein. These results suggest a model for this middle ear mucus glycoprotein, in which the native glycoprotein is a large molecular mass polymer maintained by disulphide bridges. These disulphide linked glycoprotein units are broken down into smaller units by proteolysis. The mucus glycoprotein could not be purified completely free from low molecular mass components. A glycoprotein, susceptible to proteolysis Mr 28,000-33,000 co-fractionates with the major high molecular mass mucus glycoprotein.
Department of Physiological Sciences, University of Newcastle upon Tyne, Medical School, UK.
J E
JE
FitzGerald
G G
GG
Green
F W
FW
Stafford
J P
JP
Birchall
J P
JP
Pearson
D002648:Child; D002675:Child, Preschool; D004432:Ear, Middle / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis*; D006801:Humans; D007223:Infant; D009093:Mucus / Q000032:analysis*; D010034:Otitis Media with Effusion / Q000378:metabolism*; D012394:Rosaniline Dyes; D012967:Sodium Dodecyl Sulfate; D014461:Ultracentrifugation; D014783:Viscosity
D006023:Glycoproteins; D012394:Rosaniline Dyes; D012967:Sodium Dodecyl Sulfate; C048139:Coomassie blue