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Modifying the insect cell N-glycosylation pathway with immediate early baculovirus expression vectors

Nature Biotechnology volume 14, pages 12881292 (1996) | Download Citation

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Abstract

The baculovirus-insect cell expression system is well-suited for recombinant glycoprotein production because baculovirus vectors can provide high levels of expression and insect cells can modify newly synthesized proteins in eucaryotic fashion. However, the N-glycosylation pathway of baculovirus-infected insect cells differs from the pathway found in higher eucaryotes, as indicated by the fact that glycoproteins produced in the baculovirus system typically lack complex biantennary N-linked oligosaccharide side chains containing penultimate galactose and terminal sialic acid residues. We recently developed a new type of baculovirus vector that can express foreign genes immediately after infection under the control of the viral ie 1 promoter. These immediate early baculovirus expression vectors can be used to modify the insect cell N-glycosylation pathway and produce a foreign glycoprotein with more extensively processed N-linked oligosaccharides. These vectors can also be used to study the influence of the late steps in N-linked oligosaccharide processing on glycoprotein function. Further development could lead to baculovirus-insect cell expression systems that can produce recombinant glycoproteins with complex biantennary N-linked oligosaccharides structurally identical to those produced by higher eucaryotes.

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References

  1. 1.

    and 1987. A manual of methods for bacubvirvs vectors and insect cell culture procedures. Texas Agricultural Experiment Station Bulletin No. 1555.

  2. 2.

    and 1992. Baculovirus expression vectors, pp. 265–291 in Recombinant DNA vaccines: rationale and strategies. Isaacson, R.E. (ed.). Marcel Dekker, Inc., New York.

  3. 3.

    , , and 1992. Baculovirus expression vectors: a laboratory manual. W.H. Freeman and Company, New York.

  4. 4.

    and 1985. Assembly of asparagine-linked oligosaccharides. Ann. Rev. Biochem. 54: 631–664.

  5. 5.

    and 1994. Biosynthesis and processing of the Autographa califomica nuclear polyhedrosis virus gp64 protein. Virology 205: 300–313.

  6. 6.

    , , and 1995. Purification and properties of a golgi-derived (α-1,2)-mannosidase-l from baculovirus-infected lepidopteran insect cells (IPLB-Sf21AE) with preferential activity towards mannose6-N-acetyl-glucosamine. Biochemistry 34: 2489–2495.

  7. 7.

    , , , , and 1993. Processing of asparagine-linked oligosaccharides in insect cells. N-acetylglucosaminyltrans-ferase I and II activities in cultured lepidopteran cells. Glycobiology 3: 619–625.

  8. 8.

    , , , , , and 1993. The presence of UDP-N-acetylglucosamine:α-3-D-manno-side β-1,2-N-acetylglucosaminyl transferase I activity in Spodoptera frugiperda cells (IPLB-SF-21AE) and its enhancement as a result of bacubvirus infection. J. Biol. Chem. 268: 17902–17907.

  9. 9.

    and 1995. Processing of asparagine-linked oligosaccharides in insect cells: evidence for α-mannosidase II. Glycoconjugate J. 12: 150–155.

  10. 10.

    , , , , , , et al. 1996. Isolation and characterization of a class II α-mannosidase cDNA from lepidopteran insect cells. Glycobiology. In press.

  11. 11.

    and 1981. Isolation and characterization of mosquito cell membrane glycoproteins. Biochim. Biophys. Acta 640: 655–671.

  12. 12.

    and 1984. Regulation of asparagine-linked oligosaccha-ride processing. Oligosaccharide processing in Aedes albopictus mosquito cells. J. Biol. Chem. 259: 2375–2382.

  13. 13.

    , , , and 1985. Arylphorin from Manduca sexta: carbohydrate structure and immunological studies. Arch. Biochem. Biophys. 243: 115–124.

  14. 14.

    , , and 1987. Asparagine-linked oligosaccharides of locust lipophorin. Insect Biochem. 17: 531–538.

  15. 15.

    , , , , and 1990. The oligosaccharides of influenza virus hemagglutinin expressed in insect cells by a baculovirus vector. Virology 174: 418–429.

  16. 16.

    , , and 1991. Carbohydrate variant of the recombinant β-subunit of human choriogonadotropin expressed in baculovirus expression system. J. Biol. Chem. 266: 4081–4087.

  17. 17.

    , , and 1991. Characterization of oligo-saccharide structures on a chimeric respiratory syncytial virus protein expressed in insect cell line Sf9. Biochemistry 30: 2863–2868.

  18. 18.

    , , , , , and 1991. Characterization of oligosaccharides from Drosophila melanogaster glycoproteins. Biochim. Biophys. Acta 1075: 146–153.

  19. 19.

    , , , and 1992. Determination of the glycosylation patterns, disulfide linkages, and protein heterogeneities of baculovirus-expressed mouse interleukin-3 by mass spectrometry. Biochemistry 31: 11651–11659.

  20. 20.

    , , , , and 1993. Biosynthesis and secretion of human interleukin 2 glycoprotein variants from baculovirus-infected Sf21 cells. Characterization of polypeptides and posttranslational modifications. Eur. J. Biochem. 215: 189–197.

  21. 21.

    , , , , and 1993. Site-specific N-glycosylation and Oligosaccharide structures of recombinant HIV-1 gp120 derived from a baculovirus expression system. Biochemistry 32: 11087–11099.

  22. 22.

    , , , and 1994. Structures of the N-linked oligosaccharides of the membrane glycoproteins from three lepidopteran cell lines (Sf-21, IZD-Mb-0503, Bm-N). Arch. Biochem. Biophys. 308: 148–157.

  23. 23.

    , , , , and 1994. Structural analysis and localization of the carbohydrate moieties of a soluble human interferon G receptor produced in baculovirus-infected insect cells. Protein Science 3: 30–38.

  24. 24.

    , , and 1993. Insect cell hosts for baculovirus expression vectors contain endogenous exoglycosidase activity. Biotech. Prog. 9: 146–152.

  25. 25.

    , , , , and 1995. Insect cells contain an unusual, membrane-bound β-N-acetylglu-cosaminidase probably involved in the processing of protein N-glycans. J. Biol. Chem. 270: 17344–17349.

  26. 26.

    , , , and 1996. N-acetyl-β-gluco-saminidase accounts for differences in glycosylation of influenza virus hemagglutinin expressed in insect cells from a baculovirus vector. J. Virol. 70: 4103–4109.

  27. 27.

    , , and 1996. Immediate early baculovirus vectors for foreign gene expression in transformed or infected insect cells. Protein Expression and Purification. In press.

  28. 28.

    , , and 1993. Characterization of two cis-regulatory regions in the murine β-1,4-galactosyltransferase gene: evidence for a negative regulatory element that controls initiation at the proximal site. J. Biol. Chem. 268: 14348–14359.

  29. 29.

    , , , and 1992. Beta-1,4-galac-tosyltransferase: a short NH2-terminal fragment that includes the cytoplasmic and transmembrane domain is sufficient for golgi retention. J. Biol. Chem. 267: 9241–9247.

  30. 30.

    , , and 1986. Complete sequence and enhancer function of the homologous DNA regions of Autographa californica nuclear polyhedrosis virus. J. Virol. 60: 224–229.

  31. 31.

    and 1993. The hr5 transcriptional enhancer stimulates early expression from the Autographa californica nuclear polyhedrosis virus genome but is not required for virus replication. J. Virol. 67: 5776–5785.

  32. 32.

    , , and 1983. Molecular engineering of the Autographa californica nuclear polyhedrosis virus genome: deletion mutations within the polyhedrin gene. J. Virol. 46: 584–593.

  33. 33.

    1986. The 64K envelope protein of budded Autographa californica nuclear polyhedrosis virus. Curr. Top. Microbiol. Immunol. 131: 103–118.

  34. 34.

    , and 1995. Biochemical analysis of the N-glycosylation pathway in baculovirus-infected lepidopteran insect cells. Virology 212: 500–511.

  35. 35.

    , and 1992. Baculovirus gp64 envelope glycoprotein is sufficient to mediate pH-dependent membrane fusion. J. Virol. 66: 6829–6835.

  36. 36.

    and 1985. Mechanism of neutralization of budded Autographa californica nuclear polyhedrosis virus by a monoclonal antibody: Inhibition of entry by adsorptive endocytosis. Virology 143: 185–195.

  37. 37.

    , , , and 1984. Neutralization of budded Autographa californica NPV by a monoclonal antibody: Identification of the target antigen. Virology 133: 354–362.

  38. 38.

    , , and 1989. Alteration of terminal glycosylation sequences on N-linked oligosaccharides of Chinese hamster ovary cells by expression of beta-galactoside alpha 2,6-sialyltransferase. J. Biol. Chem. 264: 13848–13855.

  39. 39.

    , , and 1995. Tissue plasminogen activator coexpressed in Chinese hamster ovary cells with alpha(2,6)-sialyltransferase contains NeuAc alpha(2,6)Gal beta(1,4)Glc-N-AcR linkages. Biotechnol. Prog. 11: 348–351.

  40. 40.

    , , , , and 1995. Construction of stable BHK-21 cells coexpressing human secretory glycoproteins and human Gal(β1-4)GlcNAc-R α 2,6-sialyltransferase: α 2,6-linked NeuAc is preferentially attached to the Gal(β1 -4)GlcNAc(β1 -2)Man(α1 -3)-branch of diantennary oligosaccharides from secreted recombinant beta-trace protein. Eur. J. Biochem. 232: 718–725.

  41. 41.

    , , , , and 1990. Transfer and expression of a murine UDP-Gal:beta-D-Gal-alpha 1,3-galactosyl-transferase gene in transfected Chinese hamster ovary cells. Competition reactions between the alpha 1,3-galactosyltransferase and the endogenous alpha 2,3-sialyltransferase. J. Biol. Chem. 265: 6225–6234.

  42. 42.

    , , , and 1995. Insertion into Aspergillus nidulans of functional UDP-GlcNAc: alpha 3-D- mannoside beta-1,2-N-acetyl-glucosaminyl-transferase I, the enzyme catalysing the first committed step from oligomannose to hybrid and complex N-glycans. Glycoconjugate J. 12: 360–370.

  43. 43.

    1993. Continuous foreign gene expression in stably-transformed insect cells, pp. 193–217 in Insect cell culture engineering. Goosen, M.F.A., Daugulis, A., and Faulkner, P. (eds.). Marcel Dekker, Inc., New York.

  44. 44.

    , , , , and 1990. Use of early baculovirus promoters for continuous expression and efficient processing of foreign gene products in stably-transformed lepidopteran cells. Bio/Technology 8: 950–955.

  45. 45.

    and 1995. Continuous foreign gene expression in transformed lepidopteran insect cells, pp. 187–202 in Methods in molecular biology Vol39: Baculovirus expression protocols. Richardson, C.D. (ed.). Humana Press, Clifton, N.J.

  46. 46.

    , , , , , and 1996. Elongation of the N-glycans of fowl plague virus hemagglutinin expressed in Spodoptera frugiperda (Sf9) cells by coexpression of human β1,2-N-acetylglucosaminyltranferase I. Glycobiology 6: 165–175.

  47. 47.

    and 1991. Asparagine-linked Oligosaccharide processing in lepidopteran insect cells. Temporal dependence of the nature of the oligosaccharides assembled on asparagine-289 of recombinant human plasminogen produced in baculovirus vector infected Spodoptera frugiperda (IPLB-SF-21AE) cells. Biochemistry 30: 6167–6174.

  48. 48.

    , , and 1990. Oligosaccharide processing in the expression of human plasminogen cDNA by lepidopteran insect (Spodoptera frugiperda) cells. Biochemistry 29: 5584–5590.

  49. 49.

    and 1988. Scaleup of insect cell cultures: Protective effects of pluronic F68. Bio/Technology 6: 1411–1418.

  50. 50.

    and 1993. A method for producing recombinant baculovirus expression vectors at high frequency. Biotechniques 14: 810–817.

  51. 51.

    , , and 1986. Galactosyltransferase-dependent sialylation of complex and endo-N-acetylglucosaminidase H-treated core N-glycans in vitro. FEBS Letters 203: 64–68.

  52. 52.

    , , , , , and 1989. Glycosyltransferase probes. Methods Enzymol. 179: 82–95.

  53. 53.

    , , , , , et al. 1985. Measurement of protein using bicinchoninic acid. Anal. Biochem. 150: 76–85.

  54. 54.

    and 1983. Monoclonal antibodies to baculovirus structural proteins: determination of specificities by Western blot analysis. Virology 125: 432–444.

  55. 55.

    and 1989. Glycosylation and secretion of human tissue plasminogen activator in recombinant baculovirus-infected insect cells. Mol. Cell. Biol. 9: 214–223.

  56. 56.

    1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.

  57. 57.

    , , , and 1984. A rapid, sensitive method for detection of alkaline phosphatase conjugated anti-antibody on western blot. Anal. Biochem. 36: 175–179.

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Author notes

    • Eric E. Finn

    Present Address: Regional Primate Research Center, University of Washington, P.O. Box 357330, Seattle, WA 98195.

    • Donald L. Jarvis

    e-mail: dljarvis@tamu.edu

Affiliations

  1. Department of Entomology and Center for Advanced Invertebrate Molecular Sciences, Texas A&M University, College Station, TX 77843.

    • Donald L. Jarvis

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DOI

https://doi.org/10.1038/nbt1096-1288

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