Isolation of high molecular weight mucins from human whole saliva by ultracentrifugation.
A high molecular weight mucin fraction was prepared from human whole saliva using relatively mild conditions. The method involves ultracentrifugation of human whole saliva in the presence of 7.2 M urea and 0.5 M sodium chloride. The resulting preparation consists of a highly purified salivary mucin fraction, as judged by several purity criteria: molecular weight analysis of the final preparation by gel electrophoresis and analytical gel filtration indicated an apparent molecular weight greater than 10(6). Analytical isopycnic density centrifugation demonstrated that the preparation consisted of a mucin fraction with a buoyant density of approximately 1.47 g/ml. The final preparation comprised 12.8% protein, 31.8% N-acetylglucosamine, 11.5% N-acetylgalactosamine, 9.5% fucose, 21.4% galactose, 0.8% mannose, 10.3% N-acetylneuraminic acid, 1.7% sulphate and 0.16% fatty acids.
Veerman
E C
EC
Department of Oral Biochemistry, Vrije Universiteit, Amsterdam.
Valentijn-Benz
M
M
Bank
R A
RA
Nieuw Amerongen
A V
AV
D002500:Centrifugation, Isopycnic; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008970:Molecular Weight; D009077:Mucins / Q000302:isolation & purification*; D012463:Saliva / Q000032:analysis*; D012965:Sodium Chloride / Q000494:pharmacology; D014508:Urea / Q000494:pharmacology
D009077:Mucins; D012965:Sodium Chloride; D014508:Urea