Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1995 Sep;69(9):5791–5797. doi: 10.1128/jvi.69.9.5791-5797.1995

The alpha-glucosidase inhibitor N-butyldeoxynojirimycin inhibits human immunodeficiency virus entry at the level of post-CD4 binding.

P B Fischer 1, M Collin 1, G B Karlsson 1, W James 1, T D Butters 1, S J Davis 1, S Gordon 1, R A Dwek 1, F M Platt 1
PMCID: PMC189444  PMID: 7543588

Abstract

The alpha-glucosidase inhibitor N-butyldeoxynojirimycin (NB-DNJ) is a potent inhibitor of human immunodeficiency virus (HIV) replication and syncytium formation in vitro. However, the exact mechanism of action of NB-DNJ remains to be determined. In this study we have examined the impairment of HIV infectivity mediated by NB-DNJ. By two independent HIV entry assays [PCR-based HIV entry assay and entry of Cocal(HIV) pseudotypes], the reduction in infectivity was found to be due to an impairment of viral entry. No effect of NB-DNJ treatment was seen on the kinetics of the interaction between gp120 and CD4 (surface plasmon resonance; BIAcore) or on the binding of virus particles to H9 cells (using radiolabeled virions). We therefore conclude that a major mechanism of action of NB-DNJ as an inhibitor of HIV replication is the impairment of viral entry at the level of post-CD4 binding, due to an effect on viral envelope components.

Full Text

The Full Text of this article is available as a PDF (505.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Block T. M., Lu X., Platt F. M., Foster G. R., Gerlich W. H., Blumberg B. S., Dwek R. A. Secretion of human hepatitis B virus is inhibited by the imino sugar N-butyldeoxynojirimycin. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2235–2239. doi: 10.1073/pnas.91.6.2235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brigham-Burke M., Edwards J. R., O'Shannessy D. J. Detection of receptor-ligand interactions using surface plasmon resonance: model studies employing the HIV-1 gp120/CD4 interaction. Anal Biochem. 1992 Aug 15;205(1):125–131. doi: 10.1016/0003-2697(92)90588-x. [DOI] [PubMed] [Google Scholar]
  3. Collin M., Gordon S. The kinetics of human immunodeficiency virus reverse transcription are slower in primary human macrophages than in a lymphoid cell line. Virology. 1994 Apr;200(1):114–120. doi: 10.1006/viro.1994.1169. [DOI] [PubMed] [Google Scholar]
  4. Collin M., Herbein G., Montaner L., Gordon S. PCR analysis of HIV1 infection of macrophages: virus entry is CD4-dependent. Res Virol. 1993 Jan-Feb;144(1):13–19. doi: 10.1016/s0923-2516(06)80006-3. [DOI] [PubMed] [Google Scholar]
  5. Collin M., James W., Gordon S. Development of techniques to analyse the formation of HIV provirus in primary human macrophages. Res Virol. 1991 Mar-Jun;142(2-3):105–112. doi: 10.1016/0923-2516(91)90045-5. [DOI] [PubMed] [Google Scholar]
  6. Datema R., Romero P. A., Rott R., Schwarz R. T. On the role of oligosaccharide trimming in the maturation of Sindbis and influenza virus. Arch Virol. 1984;81(1-2):25–39. doi: 10.1007/BF01309294. [DOI] [PubMed] [Google Scholar]
  7. Davis S. J., Ward H. A., Puklavec M. J., Willis A. C., Williams A. F., Barclay A. N. High level expression in Chinese hamster ovary cells of soluble forms of CD4 T lymphocyte glycoprotein including glycosylation variants. J Biol Chem. 1990 Jun 25;265(18):10410–10418. [PubMed] [Google Scholar]
  8. Dedera D., Vander Heyden N., Ratner L. Attenuation of HIV-1 infectivity by an inhibitor of oligosaccharide processing. AIDS Res Hum Retroviruses. 1990 Jun;6(6):785–794. doi: 10.1089/aid.1990.6.785. [DOI] [PubMed] [Google Scholar]
  9. Elbein A. D. Inhibitors of the biosynthesis and processing of N-linked oligosaccharide chains. Annu Rev Biochem. 1987;56:497–534. doi: 10.1146/annurev.bi.56.070187.002433. [DOI] [PubMed] [Google Scholar]
  10. Fleet G. W., Karpas A., Dwek R. A., Fellows L. E., Tyms A. S., Petursson S., Namgoong S. K., Ramsden N. G., Smith P. W., Son J. C. Inhibition of HIV replication by amino-sugar derivatives. FEBS Lett. 1988 Sep 12;237(1-2):128–132. doi: 10.1016/0014-5793(88)80185-6. [DOI] [PubMed] [Google Scholar]
  11. Foddy L., Hughes R. C. Assembly of asparagine-linked oligosaccharides in baby hamster kidney cells treated with castanospermine, an inhibitor of processing glucosidases. Eur J Biochem. 1988 Aug 1;175(2):291–299. doi: 10.1111/j.1432-1033.1988.tb14196.x. [DOI] [PubMed] [Google Scholar]
  12. Gregory S., Collman R., James W., Gordon S., Gonzalez-Scarano F., Nathanson N. HIV-1 pseudotype virus containing a Cocal virus genome and an HIV envelope: construction, assay and use. J Virol Methods. 1993 Oct;44(2-3):287–304. doi: 10.1016/0166-0934(93)90064-x. [DOI] [PubMed] [Google Scholar]
  13. Gross V., Andus T., Tran-Thi T. A., Schwarz R. T., Decker K., Heinrich P. C. 1-deoxynojirimycin impairs oligosaccharide processing of alpha 1-proteinase inhibitor and inhibits its secretion in primary cultures of rat hepatocytes. J Biol Chem. 1983 Oct 25;258(20):12203–12209. [PubMed] [Google Scholar]
  14. Gruters R. A., Neefjes J. J., Tersmette M., de Goede R. E., Tulp A., Huisman H. G., Miedema F., Ploegh H. L. Interference with HIV-induced syncytium formation and viral infectivity by inhibitors of trimming glucosidase. Nature. 1987 Nov 5;330(6143):74–77. doi: 10.1038/330074a0. [DOI] [PubMed] [Google Scholar]
  15. Hughes R. C., Foddy L., Bause E. Asparagine-linked oligosaccharides of BHK cells treated with inhibitors of oligosaccharide processing. Biochem J. 1987 Nov 1;247(3):537–545. doi: 10.1042/bj2470537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jones I. M., Jacob G. S. Anti-HIV drug mechanism. Nature. 1991 Jul 18;352(6332):198–198. doi: 10.1038/352198b0. [DOI] [PubMed] [Google Scholar]
  17. Karlsson G. B., Butters T. D., Dwek R. A., Platt F. M. Effects of the imino sugar N-butyldeoxynojirimycin on the N-glycosylation of recombinant gp120. J Biol Chem. 1993 Jan 5;268(1):570–576. [PubMed] [Google Scholar]
  18. Karpas A., Fleet G. W., Dwek R. A., Petursson S., Namgoong S. K., Ramsden N. G., Jacob G. S., Rademacher T. W. Aminosugar derivatives as potential anti-human immunodeficiency virus agents. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9229–9233. doi: 10.1073/pnas.85.23.9229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lubas W. A., Spiro R. G. Evaluation of the role of rat liver Golgi endo-alpha-D-mannosidase in processing N-linked oligosaccharides. J Biol Chem. 1988 Mar 15;263(8):3990–3998. [PubMed] [Google Scholar]
  20. Lubas W. A., Spiro R. G. Golgi endo-alpha-D-mannosidase from rat liver, a novel N-linked carbohydrate unit processing enzyme. J Biol Chem. 1987 Mar 15;262(8):3775–3781. [PubMed] [Google Scholar]
  21. McCune J. M., Rabin L. B., Feinberg M. B., Lieberman M., Kosek J. C., Reyes G. R., Weissman I. L. Endoproteolytic cleavage of gp160 is required for the activation of human immunodeficiency virus. Cell. 1988 Apr 8;53(1):55–67. doi: 10.1016/0092-8674(88)90487-4. [DOI] [PubMed] [Google Scholar]
  22. McKeating J. A., McKnight A., Moore J. P. Differential loss of envelope glycoprotein gp120 from virions of human immunodeficiency virus type 1 isolates: effects on infectivity and neutralization. J Virol. 1991 Feb;65(2):852–860. doi: 10.1128/jvi.65.2.852-860.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Montefiori D. C., Robinson W. E., Jr, Mitchell W. M. Role of protein N-glycosylation in pathogenesis of human immunodeficiency virus type 1. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9248–9252. doi: 10.1073/pnas.85.23.9248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Moore J. P., McKeating J. A., Jones I. M., Stephens P. E., Clements G., Thomson S., Weiss R. A. Characterization of recombinant gp120 and gp160 from HIV-1: binding to monoclonal antibodies and soluble CD4. AIDS. 1990 Apr;4(4):307–315. doi: 10.1097/00002030-199004000-00004. [DOI] [PubMed] [Google Scholar]
  25. Moore J. P., McKeating J. A., Weiss R. A., Sattentau Q. J. Dissociation of gp120 from HIV-1 virions induced by soluble CD4. Science. 1990 Nov 23;250(4984):1139–1142. doi: 10.1126/science.2251501. [DOI] [PubMed] [Google Scholar]
  26. Moore S. E., Spiro R. G. Demonstration that Golgi endo-alpha-D-mannosidase provides a glucosidase-independent pathway for the formation of complex N-linked oligosaccharides of glycoproteins. J Biol Chem. 1990 Aug 5;265(22):13104–13112. [PubMed] [Google Scholar]
  27. Pal R., Hoke G. M., Sarngadharan M. G. Role of oligosaccharides in the processing and maturation of envelope glycoproteins of human immunodeficiency virus type 1. Proc Natl Acad Sci U S A. 1989 May;86(9):3384–3388. doi: 10.1073/pnas.86.9.3384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pinter A., Honnen W. J. Biochemical characterization of cell-associated and extracellular products of the Friend spleen focus-forming virus env gene. Virology. 1989 Nov;173(1):136–143. doi: 10.1016/0042-6822(89)90229-8. [DOI] [PubMed] [Google Scholar]
  29. Pinter A., Honnen W. J., Li J. S. Studies with inhibitors of oligosaccharide processing indicate a functional role for complex sugars in the transport and proteolysis of Friend mink cell focus-inducing murine leukemia virus envelope proteins. Virology. 1984 Jul 15;136(1):196–210. doi: 10.1016/0042-6822(84)90259-9. [DOI] [PubMed] [Google Scholar]
  30. Rabouille C., Spiro R. G. Nonselective utilization of the endomannosidase pathway for processing glycoproteins by human hepatoma (HepG2) cells. J Biol Chem. 1992 Jun 5;267(16):11573–11578. [PubMed] [Google Scholar]
  31. Ratner L., Vander Heyden N. Mechanism of action of N-butyl deoxynojirimycin in inhibiting HIV-1 infection and activity in combination with nucleoside analogs. AIDS Res Hum Retroviruses. 1993 Apr;9(4):291–297. doi: 10.1089/aid.1993.9.291. [DOI] [PubMed] [Google Scholar]
  32. Ratner L., vander Heyden N., Dedera D. Inhibition of HIV and SIV infectivity by blockade of alpha-glucosidase activity. Virology. 1991 Mar;181(1):180–192. doi: 10.1016/0042-6822(91)90483-r. [DOI] [PubMed] [Google Scholar]
  33. Repp R., Tamura T., Boschek C. B., Wege H., Schwarz R. T., Niemann H. The effects of processing inhibitors of N-linked oligosaccharides on the intracellular migration of glycoprotein E2 of mouse hepatitis virus and the maturation of coronavirus particles. J Biol Chem. 1985 Dec 15;260(29):15873–15879. doi: 10.1016/S0021-9258(17)36339-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Saunier B., Kilker R. D., Jr, Tkacz J. S., Quaroni A., Herscovics A. Inhibition of N-linked complex oligosaccharide formation by 1-deoxynojirimycin, an inhibitor of processing glucosidases. J Biol Chem. 1982 Dec 10;257(23):14155–14161. [PubMed] [Google Scholar]
  35. Schlesinger S., Koyama A. H., Malfer C., Gee S. L., Schlesinger M. J. The effects of inhibitors of glucosidase I on the formation of Sindbis virus. Virus Res. 1985 Mar;2(2):139–149. doi: 10.1016/0168-1702(85)90244-8. [DOI] [PubMed] [Google Scholar]
  36. Simon J. H., Somoza C., Schockmel G. A., Collin M., Davis S. J., Williams A. F., James W. A rat CD4 mutant containing the gp120-binding site mediates human immunodeficiency virus type 1 infection. J Exp Med. 1993 Apr 1;177(4):949–954. doi: 10.1084/jem.177.4.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tan A., van den Broek L., van Boeckel S., Ploegh H., Bolscher J. Chemical modification of the glucosidase inhibitor 1-deoxynojirimycin. Structure-activity relationships. J Biol Chem. 1991 Aug 5;266(22):14504–14510. [PubMed] [Google Scholar]
  38. Taylor D. L., Kang M. S., Brennan T. M., Bridges C. G., Sunkara P. S., Tyms A. S. Inhibition of alpha-glucosidase I of the glycoprotein-processing enzymes by 6-O-butanoyl castanospermine (MDL 28,574) and its consequences in human immunodeficiency virus-infected T cells. Antimicrob Agents Chemother. 1994 Aug;38(8):1780–1787. doi: 10.1128/aac.38.8.1780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Walker B. D., Kowalski M., Goh W. C., Kozarsky K., Krieger M., Rosen C., Rohrschneider L., Haseltine W. A., Sodroski J. Inhibition of human immunodeficiency virus syncytium formation and virus replication by castanospermine. Proc Natl Acad Sci U S A. 1987 Nov;84(22):8120–8124. doi: 10.1073/pnas.84.22.8120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Willey R. L., Bonifacino J. S., Potts B. J., Martin M. A., Klausner R. D. Biosynthesis, cleavage, and degradation of the human immunodeficiency virus 1 envelope glycoprotein gp160. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9580–9584. doi: 10.1073/pnas.85.24.9580. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES