Abstract
Glycoproteins commercially available in multi-gram quantities, were used to prepare milligram amounts of neoglycoproteins. The glycoproteins bromelain and bovine γ-globulin were proteolyzed to obtain glycopeptides or converted to a mixture of glycans through hydrazinolysis. The glycan mixture was structurally simplified by carbohydrate remodeling using exoglycosidases. Glycopeptides were biotinylated using N-hydroxysuccinimide activated-long chain biotin while glycoprotein-derived glycans were first reductively aminated with ammonium bicarbonate and then biotinylated. The resulting biotinylated carbohydrates were structurally characterized and then bound to streptavidin to afford neoglycoproteins. The peptidoglycan component of raw, unbleached heparin (an intermediate in the manufacture of heparin) was similarly biotinylated and bound to streptavidin to obtain milligram amounts of a heparin neoproteoglycan. The neoglycoconjugates prepared contain well defined glycan chains at specific locations on the streptavidin core and should be useful for the study of protein-carbohydrate interactions and affinity separations.
Similar content being viewed by others
References
Kobata A (1998) Glycoconjugate J 15: 323–31.
Jenkins N, Parekh RB, James DC (1996) Nature Biotechnol 14:975–81.
Hounsell DF, Renouf EV (1998) Adv Exper Med & Biol 435:251–60.
Sears P, Wong CH (1998) Cellular & Molec Life Sci 54: 223–52.
Lee KB, Loganathan D, Merchant ZM, Linhardt RJ (1990) Appl Biochem Biotechnol 23: 53–80.
Iozzo RV (1998) Ann Rev Biochem 67: 609–52.
Moens S, Vanderleyden J (1997) Arch Microbiol 168: 169–75.
Wolff MW, Murhammer DW, Linhardt RJ (1999) Prep Biochem 29: 1–21.
Nelson RM, Venot A, Bevilacqua MP, Linhardt RJ, Stamenkovic I (1995) Annu Rev Cell Dev Biol 11: 601–31
Choe J, VanderNoot VA, Linhardt RJ, Dordick JS (1998) AlChE J 44: 2542–8.
Quesenberry MS, Lee RT, Lee YC(1997) Biochemistry 36: 2724–32
Wong SYC (1995) Curr Opin Struct Biol 5: 599–604.
Kawaguchi K, Kuhlenschmidt M, Roseman S, Lee YC (1981) J Biol Chem 265: 2230–4.
Garcia-Cascado G, Sanchez-Monge R, Chrispeels MJ, Armentia A, Salcedo G, Gomez L (1996) Glycobiology 6: 471–7.
Nadkarni VD, Linhardt RJ (1996) Biotechniques 23: 382–5.
Caldwell EEO, Andreasen AM, Blietz MA, Serrahn JN, Vander-Noot V, Park Y, Yu G, Linhardt RJ, Weiler JM (1999) Arch Biochem Biophys 361: 215–22.
Shinohara T, Sota, H, Fimi K, Shimizu M, Gotoh M, Tosu M, Hasegawa Y (1995) J Biochem 117: 1076–82.
Chen VJ, Wold F (1986) Biochemistry 25: 939–44.
Bay S, Berthier-Vergnes O, Cantacuzene D (1997) Carbohydr Res 298: 153–61.
Hasegawa Y, Shinohara Y, Sota H, Shimizu M, Vretblad P, Bhik-habhai R, Fagerstam LG, Ekstrom B, Blikstad I (1996) Chemistry Lett 40–2.
Shinohara Y, Sota H, Gotoh M, Hasebe M, Tosu M, Nakao J, Hasegawa Y (1996) Anal Chem 68: 2573–9.
Toomre DK, Varki A (1994) Glycobiology 4: 653–63.
Wong SYC, Manger ID, Guile GR, Rademacher TW, Dwek RA (1993) Biochem J 296: 817–25.
Leteux C, Childs RA, Wengang C, Stoll MS, Kogelberg H and Feizi T, (1998) Glycobiology 8: 227–36.
Gerber SA, Scott R, Turecek F, Gelb MH (1999) J Am Chem Soc 121: 1102–3.
Cummings RD (1994) In Methods in Enzymology, vol 230 (Lennarz WJ, Hart GW, eds) pp 66–86. San Diego: Academic Press.
Al-Hakim A, Linhardt RJ (1991) Appl Theoret Electrophor 1:305–12.
Fedarko NS (1993) Experientia 49: 369–83.
Chandrasekhar S, Estermar MA, Hoffman HA (1987) Anal Biochem 161: 103–8.
Lommerse JPM, Kroon-Batenburg LMJ, Kamerling, JP and Vliegenthart JFG (1995) Biochemistry 34: 8196–206.
Fujii S, Nishiura T, Nishikawa A, Miura R, Taniguchi N (1990) J Biol Chem 265: 6009–18.
Lee YC, Lee RT (1994) Neoglycoconjugates: Preparation and Application.San Diego: Academic Press.
Linhardt RJ (1991) Chem & Indust 2: 45–50.
Liu J, Desai UR, Han X, Toida T, Linhardt RJ (1995) Glycobiology 765–74.
Ripellino JA, Margalis RU (1989) Neurochem 52: 807–12.
Schneller M, Geiger RE (1992) Biol Chem Hoppe-Seyler 373:1095–104.
Linhardt RJ, Toida T (1997) In Carbohydrates as Drugs (Witczak ZB, Nieforth KA, eds) pp 277–341. New York: Marcell Dekker.
Masson L, Mazza A, Brousseau R (1994) Anal Biochem 218:405–12.
Wadhwa MS, Rice KG (1995) J Drug Targeting 3: 111–27.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kuberan, B., Gunay, N., Dordick, J. et al. Preparation and isolation of neoglycoconjugates using biotin-streptavidin complexes. Glycoconj J 16, 271–281 (1999). https://doi.org/10.1023/A:1007009927087
Issue Date:
DOI: https://doi.org/10.1023/A:1007009927087
- neoglycoconjugates
- neoglycoprotein
- neoproteoglycan
- biotin
- streptavidin
- heparin
- bromelain
- γ-globulin
- MWCO, molecular weight cut-off
- ANTS, 8-amino-1,3,6-napthalenetrisulfonate
- DMSO,dimethyl sulfoxide
- ESI,electrospray ionization
- NHS,N-hydroxysuccinimide
- LC,long chain
- CE,capillary electrophoresis
- PAGE,polyacryamide gel electrophoresis
- SDS,sodium dodecylsulfate
- BSA,bovine serum albumin
- SAX,strong-anion exchange