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Keywords:

  • bovine milk;
  • proteose peptone;
  • component PP3;
  • glycoprotein;
  • glycan

Abstract

  1. Top of page
  2. Abstract
  3. References

The heat-stable acid-soluble phosphoglycoprotein component PP3 was isolated from the bovine milk proteose peptone fraction by concanavalin A affinity chromatography. Glycopeptides were released by pronase digestion of the milk component PP3 and were subsequently separated by high-pH anion-ex-change chromatography on Carbopac PA-1. The primary structures of the glycan and peptide moieties of eight N-glycopeptides have been established by combining methylation analysis, mass spectrometry, 400-MHz 1H-NMR spectroscopy, and peptide sequence analysis. All the analyzed fractions contained biantennary N-acetyllactosamine-type carbohydrate chains, some of them with a GalNAc(β1–4)GlcNAc or a NeuAc(α2–6)GalNAc(β1–4)GlcNAc group. This particular sequence did or did not replace the Gal(β–4)GlcNAc group usually found in most N-linked glycans. Moreover, the sialylated Gal and GalNAc residues were only found on the Man(α1–3) antenna.

Abbreviations
ConA

concanavalin A

CAM

cell adhesion molecule

GP

glycopeptide

HPAE

high-pH anion exchange

MFGM

milk fat globule membrane

PAD

pulsed amperometric detection

PP

proteose peptone

Enzyme
 

Mycolysin (pronase) (EC 3.4.24.31)

References

  1. Top of page
  2. Abstract
  3. References
  • Allen, G. (1981) Determination of peptide sequences, in Laboratory techniques in biochemistry and molecular biology: sequencing of proteins and peptides (Work, T. S. & Burdon, R. H., eds) pp. 178180, Elsevier/North-Holland Biochemical Press, Amsterdam .
  • Bergwerff, A. A., Thomas-Oates, J. E., Van Oostrum, J., Kamerling, J. P. & Vliegenthart, J. F. G. (1992) Human urokinase contains GalNAcβ(1–4)[Fucα(1–3)]GlcNAcβ(1–2) as a novel terminal element in N-linked carbohydrate chains, FEBS Lett. 314, 319394.
  • Bergwerff, A. A., Van Oostrum, J., Kamerling, J. P. & Vliegenthart, J. F. G. (1995) The major N-linked carbohydrate chains from human urokinase. The occurrence of 4–O-sulfated, (α2–6)-sialylated or (α1–3)-fucosylated N-acetylgalactosamine (β1–4)-N-acetylglucosamine elements, Eur J. Biochem. 228, 10091019.
  • Ciucanu, I. & Kerek, F. (1984) A simple and rapid method for the permethylation of carbohydrates, Carbohydr. Res. 131, 209217.
  • Coddeville, B., Strecker, G., Wieruszeski, J. M., Vliegenthart, J. F. G., Egge, H. & Spik, G. (1992) Heterogeneity of bovine lactotransferrin glycans. Characterization of α-d-Galp-(1–3)-β-d-Gal- and α-NeuAc-(2–6)-β-d-GalpNAc-(1–4)-β-d- GlcNAc-substiuted N-linked glycans, Carbohydr. Res. 236, 145164.
  • Courthaudon, J. L., Girardet, J. M., Chapal, C., Lorient, D. & Linden, G. (1995) Surface activity and competitive adsorption of milk component-3 and porcine pancreatic lipase at the dodecane-water interface, in Food macromolecules and colloids (Dickinson, E., ed.) pp. 5870, Royal Society of Chemistry, London .
  • Fournet, B., Strecker, G., Leroy, Y. & Montreuil, J. (1981) Gas-liquid chromatography and mass spectrometry of methylated and acetylated methyl glycosides. Application to the structural analysis of glycoprotein glycans, Anal. Biochem. 116, 489502.
  • Girardet, J. M., Linden, G., Loye, S., Courthaudon, J. L. & Lorient, D. (1993) Study of mechanism of lipolysis inhibition by bovine milk proteose peptone component-3, J. Dairy Sci 76, 21562163.
  • Girardet, J. M., Saulnier, F., Driou, A., Linden, G., Coddeville, B. & Spik, G. (1994) High performance electrophoresis chromatography of bovine milk component-3 glycoproteins, J. Dairy Sci. 77, 12051216.
  • Green, E. D., van Halbeek, H., Boime, I. & Baezinger, J. U. (1985) Structural elucidation of the disulfated oligosaccharide from bovine lutropin, J. Biol. Chem. 260, 1562315630.
  • Hardy, M. R. & Townsend, R. R. (1988) Separation of positional isomers of oligosaccharides and glycopeptides by high-performance anion-exchange chromatography with pulsed amperometric detection, Proc. Natl Acad. Sci. USA 85, 32893293.
  • Johnsen, L. B., Sørensen, E. S., Petersen, T. E. & Berglund, L. (1995) Characterization of a bovine, mammary gland PP3 cDNA reveals homology with a mouse and rat immume, protein glycosylation-dependent CAM-1, Biochim. Biophys. Acta 1260, 116118.
  • Kamerling, J. P., Gerwig, G. J., Vliegenthart, J. F. G. & Clamp, J. R. (1975) Characterization by gas liquid chromatography mass spectrometry of pertrimethylsilyl glycosides obtained in the methanolysis of glycoproteins and glycosides, Biochem. J. 151, 491495.
  • Kanno, C. (1989) Purification and separation of multiple forms of lactophorin from bovine milk whey and their immunological and electrophoretic properties, J. Dairy Sci. 72, 883891.
  • Kanno, C. & Othake, Y. (1981) Immunological and electrophorectic characteristics of anti-SGP reacting protein among component-3,-5 and-8 fractions of proteose peptone of bovine milk: evidence for the multiple forms, Jpn. J. Zootech. Sci. 52, 282296.
  • Kester, J. J. & Brunner, J. R. (1982) Milk fat-globule membrane as possible origin of proteose peptone glycoproteins, J. Dairy Sci. 65, 22412252.
  • Krusius, T., Finne, J. & Ravaula, H. (1976) The structural basis of the different affinities of two types of acidic N-glycosidic glycopeptides for concanvalin A-Sepharose, FEBS Lett, 71, 117120.
  • Lasky, I. A., Singer, M. S., Dowbenko, D., Imai, Y., Henzel, W. J., Grimley, C., Fennie, C., Gillett, N., Watson, S. R. & Rosen, S. D. (1992) An endothelial ligand for L-selection is a novel mucin-like molecule, Cell 69, 927938.
  • Mengeling, B. J., Manzella, S. M. & Baenziger, J. U. (1995) A cluster of basic amino acids within an α-helix is essential for α-subunit recognition by the glycoprotein hormone N-acetylgalactosaminyltransferase, Proc. Natl Acad. Sci. USA 92, 502506.
  • Montreuil, J. (1980) Primary structure of glycoprotein glycans: basis for the molecular biology of glycoproteins. Adv. Carbohydr. Chem. Biochem. 37, 157223.
  • Nakata, N., Furukawa, K., Greenwalt, D. E., Sato, T., & Kobata, A. (1993) Structural study of the sugar chains of CD36 purified from bovine mammary epithelial cells: occurrence of novel hybrid-type sugar chains containing the NeuAcα2-6GalNAcβ1–4GlcNAc and the Manα1–2Manα1–3Manα1–6Man groups. Biochemistry 32, 43694383.
  • Neeleman, A. P., Van Der Knaap, W. P. W. & Van Den Eijnden, D. H. (1994) Identification and characterization of a UDP-GalNAc:Glc-NAcβ-R β1–4-N-acetylgalactosaminyltransferase from cercariae of the schistosome Trichobilharzia ocellata. Catalysis of a key step in the synthesis of N,N′-diacetyllactosediamino (lacdiNAc)-type glycans. Glycobiology 4, 641651.
  • Nejjaar, Y., Pâquet, D., Godbillon, G. & Le Deaut, J. Y. (1986) Immunological relationship between the hydrophobic fraction of proteose peptone and the milk fat globule membrane of bovine milk, Int. J. Biochem. 18, 893900.
  • Pâquet, D. (1989) Review: the proteose peptone fraction of milk, Lait 69, 121.
  • Pâquet, D., Nejjar, Y. & Linden, G. (1988) Study of a hydrophobic protein fraction isolated from milk proteose peptone, J. Dariy Sci. 71, 14641471.
  • Sato, T., Furukawa, K., Greenwalt, D. E. & Kobata, A. (1993) Most bovine milk fat globule membrane glycoproteins contain asparagine-linked sugar chains with GalNAcβ1–4GlcNAc groups, J. Biochem. (Tokyo) 114, 890900.
  • Sato, T., Takio, K., Kobata, A., Greenwalt, D. E. & Furukawa, K. (1995) Site-specific g1ycosylation of bovine butyrophilin, J. Biochem. (Tokya) 117, 147157.
  • Smith, P. L. & Baenziger, J. U. (1992) Molecular basis of recognition hormone-specific N-acetylgalactosamine-transferase, Proc. Natl Acad. Sci. USA 89, 329333.
  • Sørensen, E. S. & Petersen, T. F., (1993) Phosphorylation, glycosylation and amino acid sequence of component PP3 from the proteose peptone fraction of bovine milk, J. Dairy Res. 60, 535542.
  • Van Halbeek, H. (1993) Structural characterization of the carbohydrate moieties of glycoproteins by high-resolution 1H-NMR spectroscopy, in Methods in molecular biology (Jones, C., Mulloy, B. & Thomas, A. H.) vol. 17, pp. 115118, Humana Press, Totowa .
  • Vliegenthart, J. F. G., Dorland, L. & van Halbeek, H. (1983) High-resolution, 1H-nuclear magnetic resonance spectroscopy as a tool in the structural analysis of carbohydrates related to glycoproteins, Adv. Carbohydr. Chem. Biochem. 41, 209374.
  • Weisshaar, G., Hiyama, J. & Renwick, A. G. C. (1990) Site specific N-glycosylation of ovine lutropin; structural analysis by one- and two-dimensional 1H-NMR spectroscopy, Eur. J. Biochem. 192, 741751.
  • Weisshaar, G., Hiyama, J., Renwick, A. G. C. & Nimtz, M. (1991) NMR investigations of the N-linked oligosaccharides at individual glycosylation sites of human lutropin, Eur. J. Biochem. 195, 257268.
  • Yan, S. B., Chao, Y. B. & van Halbeek, H. (1993) Novel Asn-linked oligosaccharides terminating in GalNAcβ(1–4)[Fucα(1 – 3)] GlcNAcβ(1–.) are present in recombinant human protein C expressed in human kidney 293 cells, Glycobiology 3, 597608.
  • Zanetta, J. P., Breckenridge, W. C. & Vincendon, G. (1972) Analysis of monosaccharides by gas-liquid chromatography of the O-methyl glycosides as trifluoroacetates derivatives. Application to glycoproteins and glycolipids, J. Chromatogr. 69, 291304.