Purification of mucin glycoproteins by density gradient centrifugation in cesium trifluoroacetate.
A procedure for the rapid isolation of mucin glycoprotein by density gradient centrifugation in cesium trifluoroacetate (CsTFA) is described. The separation of mixtures of rat tracheobronchial mucin, DNA, hyaluronic acid, and bovine serum albumin in CsTFA gradients was superior to that in cesium bromide gradients. Inclusion of guanidinium chloride or urea in the gradient had no influence on the separation obtained. The mucins isolated from sputum samples of cystic fibrosis patients by this procedure are largely free of nucleic acid, nonglycosylated proteins, and glycosaminoglycans. The results of the use of CsTFA gradient centrifugation for the isolation of mucin from extracts of bovine submaxillary gland are also presented. The CsTFA method is particularly suitable for the high-yield isolation of mucin from individual samples which are available in limited quantities.
Devaraj
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Department of Biological Chemistry and the Cell and Molecular Biology Program, Milton S. Hershey Medical Center, Pennsylvania State University, Hershey 17033.
Devaraj
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Bhavanandan
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VP
D000818:Animals; D001965:Bromides; D001980:Bronchi / Q000378:metabolism; D002417:Cattle; D002499:Centrifugation, Density Gradient / Q000379:methods; D002586:Cesium; D003550:Cystic Fibrosis / Q000503:physiopathology; D004247:DNA / Q000032:analysis; D019791:Guanidine; D006146:Guanidines; D006801:Humans; D006820:Hyaluronic Acid / Q000032:analysis; D009077:Mucins / Q000302:isolation & purification*; D011489:Protein Denaturation; D011506:Proteins / Q000032:analysis; D051381:Rats; D013183:Sputum / Q000737:chemistry; D013363:Submandibular Gland / Q000737:chemistry; D014020:Tissue Extracts / Q000737:chemistry; D014132:Trachea / Q000378:metabolism; D014269:Trifluoroacetic Acid
D001965:Bromides; D006146:Guanidines; D009077:Mucins; D011506:Proteins; D014020:Tissue Extracts; C078556:cesium bromide; D002586:Cesium; D006820:Hyaluronic Acid; D004247:DNA; D014269:Trifluoroacetic Acid; D019791:Guanidine