Abstract
Two mucins were isolated from bovine submandibular glands and termed major and minor on a quantitative basis. The major mucin representing over 80% of the total glycoprotein fraction contained 37% of its dry weight as protein in contrast to 62% for the minor mucin. Differences in the amino acid composition reflected the higher proportion of typically non-glycosylated peptide in the minor mucin. The molar ratio ofN-acetylgalactosamine to serine plus threonine was 0.82 in major and 0.65 in minor mucins, indicating a lower degree of substitution of potential glycosylation sites in the minor mucin.
Differences in the carbohydrate composition were found largely related to the sialic acids, with higher relative amounts ofN-glycoloylneuraminic acid in the minor mucin. In addition, the proportion of di-O-acetylated sialic acids was higher in the major mucin. The rate of sialidase action on the two mucins could be correlated with the content ofN-glycoloylneuraminic acid in each glycoprotein. There was no difference in the type of oligosaccharide found in each mucin and the differences in relative proportions reflected the monosaccharide composition for the two mucins. Gel filtration on Sepharose CL 2B showed a lower molecular weight distribution for the minor in contrast to the major mucin which was partially excluded. Density gradient centrifugation reflected this variation. SDS-PAGE demonstrated a regular banding pattern for the major mucin with a lowest subunit size of 1.8×105 Da and aggregates in excess of 106 Da, while the minor mucin ranged from 3.0 × 105 to 106 Da. The chemical composition of the isolated mucins was compared with previous histochemical analysis of mucin distribution in bovine submandibular glands and indicates a possible cellular location for each mucin.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- PBS:
-
0.01m sodium phosphate buffer, pH 7.3, containing 0.15m NaCl
- Neu5Ac:
-
N-acetylneuraminic acid
- Neu5Gc:
-
N-glycoloylneuraminic acid
- GalNAc-ol:
-
N-acetylgalactosaminitol
References
Veh RW, Corfield AP, Schauer R, Andres KH (1979) InProc Vth Int Symp Glycoconjugates (Schauer R, Boer P, Buddecke E, Kramer MF, Vliegenthart JFG, Wiegandt H, eds) pp 191–94. Stuttgart: Thieme.
Schauer R, Casals-Stenzel J, Corfield AP, Veh RW (1988)Glycoconjugate J 5:257–70.
Tettamanti G, Pigman W (1968)Arch Biochem Biophys 124:41–50.
Bertolini M, Pigman W (1970)Carbohydr Res 14:53–63.
Tsuji T, Osawa T (1986)Carbohydr Res 151:391–402.
D'Arcy SM, Donoghue CM, Koeleman CAM, van den Eijnden DH, Savage AV (1989)Biochem J 260:389–93.
Reuter G. Pfeil R, Stoll S, Schauer R, Kamerling JP, Versluis C, Vliegenthart JFG (1983)Eur J Biochem 134:139–43.
Bhavanandan VP, Hegarty JD (1987)J Biol Chem 262:5913–17.
Bhargava AK, Woitach JT, Davidson EA, Bhavanandan VP (1990)Proc Natl Acad Sci U S A 87:6798–802.
Corfield AP, Wember M, Schauer R, Rott R (1982)Eur. J. Biochem. 124:521–25.
Spiro RG, Bhoyroo VD (1974)J Biol Chem 249:5704–17.
Rana SS, Chandrasekaran EV, Kennedy J, Mendicino J (1984)J Biol Chem 259:12899–907.
Bayard B, Kerckaert JP, Roux D, Strecker G (1979) InProtides of Biological Fluids (Peeters H, ed.) pp 153–56. Oxford: Pergamon Press.
Clamp JR, Fraser G, Read AE (1981)Clin Sci 61:229–34.
Veh RW, Corfield AP, Sander M, Schauer R (1977)Biochim Biophys Acta 486:145–60.
Clamp JR, Bhatti T, Chambers RE (1971)Methods Biochem Anal 19:229–344.
Schauer R (1978)Methods Enzymol 50:64–89.
Schauer R (1987)Methods Enzymol 138:132–61.
Haverkamp J, Kamerling JP, Vliegenthart JFG, Veh RW, Schauer R (1977)FEBS Lett 73:215–19.
Clamp JR, Cooper B, Creeth JM, Ene D, Barrett J, Gough M (1983)Biochem J 215:421–23.
Corfield AP, Veh RW, Wember M, Michalski J-C, Schauer R (1981)Biochem J 197:293–99.
Corfield AP, Sander-Wewer M, Veh RW, Wember M, Schauer R (1986)Biol Chem Hoppe-Seyler 367:433–39.
Carlstedt I, Lindgren H, Sheehan JK, Ulmsten U, Wingerup L (1983)Biochem J 211:13–22.
Thornton DJ, Holmes DF, Sheehan JK, Carlstedt I (1989)Anal Biochem 182:160–4.
Laemmli UK (1970)Nature 227:680–5.
Sarris AH, Palade GE (1979)J Biol Chem 254:6724–31.
Mantle M, Allen A (1978)Biochem Soc Trans 6:607–9.
Mentlein R, Heiland S, Heymann E (1980)Arch Biochem Biophys 200:547–59.
Clarke AE, Denborough MA (1972)Biochem J 121:891–16.
Ames BN (1966)Methods Enzymol 8:115–18.
Schauer R, Faillard H (1968)Hoppe-Seylers Z Physiol Chem 349:961–8.
Veh RW, Meesen D, Kunz HD, May B (1982) InColonic Carcinogenesis (Malt RA, Williamson RCN, eds), pp 355–65. Lancaster: MTP Press.
Roe R, Corfield AP, Williamson RCN (1989)Histochem J 21:216–22.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Corfield, A.P., Do Amaral Corfield, C., Veh, R.W. et al. Characterization of the major and minor mucus glycoproteins from bovine submandibular gland. Glycoconjugate J 8, 330–339 (1991). https://doi.org/10.1007/BF00731345
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00731345