Characterization of gastric mucoproteins isolated by equilibrium density-gradient centrifugation in caesium chloride.
1. The mucoprotein from pig gastric mucus has been purified by equilibrium centrifugation in a CsCl gradient. 2. This procedure removes the non-covalently bound protein, which is closely associated with the mucoprotein and not easily removed from it by gel filtration. 3. The purified mucoprotein is separable by gel filtration into a high-molecular-weight mucoprotein A (mol.wt. 2.3x10(6)) and a low-molecular-weight mucoprotein B/C (mol.wt. 1.15x10(6)). 4. These two mucoproteins have the same chemical analysis namely fucose 11.3%, galactose 26%, glucosamine 19.5%, galactosamine 8.3% and protein 13.6%. 5. Mucoprotein A contains 3.1% ester sulphate. 6. These mucoproteins are isolated without enzymic digestion and have a higher protein content than the blood-group-substance mucoproteins from proteolytic digestion of gastric mucus. Detailed amino acid analysis shows that the extra protein in the non-enzymically digested material is composed of amino acids other than serine and threonine. 7. Mucoproteins A and B/C contain respectively 130 and 9 half-cystine residues per molecule of which about 78 and 6 residues are involved in disulphide linkages. 8. Cleavage of these disulphide linkages by mercaptoethanol splits both mucoproteins into four equally sized subunits of mol.wt. 5.2x10(5) for mucoprotein A and 2.8x10(4) for mucoprotein B/C. 9. The sole N-terminal amino acid of mucoprotein A is aspartic acid, whereas mucoprotein B/C has several different N-terminal amino acid residues.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001224:Aspartic Acid / Q000032:analysis; D002499:Centrifugation, Density Gradient; D003553:Cystine / Q000032:analysis; D004220:Disulfides; D004952:Esters / Q000032:analysis; D005643:Fucose / Q000032:analysis; D005688:Galactosamine / Q000032:analysis; D005690:Galactose / Q000032:analysis; D005753:Gastric Mucosa / Q000032:analysis*; D005944:Glucosamine / Q000032:analysis; D008623:Mercaptoethanol; D008970:Molecular Weight; D009088:Mucoproteins / Q000032:analysis / Q000302:isolation & purification*; D011506:Proteins / Q000032:analysis; D013431:Sulfates / Q000032:analysis; D013552:Swine
D000596:Amino Acids; D004220:Disulfides; D004952:Esters; D009088:Mucoproteins; D011506:Proteins; D013431:Sulfates; D005643:Fucose; D001224:Aspartic Acid; D003553:Cystine; D008623:Mercaptoethanol; D005688:Galactosamine; D005944:Glucosamine; D005690:Galactose