Isolation and partial characterization of serine- and threonine-rich porcine gastric mucus glycopeptides.
1. Two subfractions from purified porcine gastric mucus glycopeptide were found to separate from each other by cesium chloride equilibrium centrifugation. The highest density fraction and two lower density fractions separated were designated VHD, HD and LD, respectively. A comparative study of these components was made. 2. The high and low density fractions, HD and LD, appeared almost the same or identical, while VHD differed completely from either of them in the following respects: (1) VHD exhibited strong alcian blue binding activity. (2) 57% of VHD bound to the DEAE-Toyopearl column equilibrated with 0.2 M NaCl. (3) VHD eluted from the Sephacryl S-400 column as a lower molecular subunit. (4) One third of the sialic acid as a minor component in VHD was constituted by N-glycolylneuraminic acid. (5) Carbohydrate composition showed typical mucus glycoprotein with slightly higher fucose content. (6) Amino acid compositions of the anionic components prepared from VHD showed the highest Ser/Thr ratio, 1.92 compared to 0.46 for LD and 0.62 for HD. (7) Oligosaccharide released from VHD by alkaline-sodium borohydride treatment was larger than that from HD or LD. 3. The above results indicate the minor component, VHD, separated from the major components, to be a quite similar but not identical component to the so-called sulfated mucus glycoprotein reported previously [Slomiany et al. (1972) J. biol. Chem. 247, 5062-5070].
Iwase
H
H
Department of Biochemistry, School of Medicine, Kitasato University, Kanagawa, Japan.
Ishii-Karakasa
I
I
Hotta
K
K
D000423:Alcian Blue; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002499:Centrifugation, Density Gradient; D002853:Chromatography, Liquid / Q000379:methods; D004588:Electrophoresis, Cellulose Acetate; D005753:Gastric Mucosa / Q000737:chemistry*; D006020:Glycopeptides / Q000737:chemistry / Q000302:isolation & purification*; D009093:Mucus / Q000737:chemistry*; D009844:Oligosaccharides / Q000737:chemistry; D010434:Pepsin A; D012694:Serine / Q000032:analysis*; D013552:Swine / Q000378:metabolism*; D013912:Threonine / Q000032:analysis*
D000596:Amino Acids; D002241:Carbohydrates; D006020:Glycopeptides; D009844:Oligosaccharides; D013912:Threonine; D012694:Serine; D010434:Pepsin A; D000423:Alcian Blue