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. 1995 Oct;69(10):6557–6562. doi: 10.1128/jvi.69.10.6557-6562.1995

Binding kinetics of ecotropic (Moloney) murine leukemia retrovirus with NIH 3T3 cells.

H Yu 1, N Soong 1, W F Anderson 1
PMCID: PMC189559  PMID: 7666559

Abstract

A quantitative analysis of the binding kinetics of intact Moloney murine leukemia retrovirus (MoMuLV) particles with NIH 3T3 cells was performed with an immunofluorescence flow cytometry assay. The virus-cell binding equilibrium dissociation constant (KD), expressed in terms of virus particle concentration, was measured to be 8.5 (+/- 6.4) x 10(-12) M at 4 degrees C and was three- to sixfold lower at temperatures above 15 degrees C. The KD of virus binding is about 1,000-fold lower than the KD of purified MoMuLV envelope. The association rate constant was determined to be 2.5 (+/- 0.9) x 10(9) M-1 min-1 at 4 degrees C and was 5- to 10-fold higher at temperatures above 15 degrees C. The apparent dissociation rate constant at 4 degrees C was 1.1 (+/- 0.4) x 10(-3) min-1 and was doubled for every 10 degrees C increase in temperature over the range tested (15 to 37 degrees C).

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Selected References

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  1. Albritton L. M., Kim J. W., Tseng L., Cunningham J. M. Envelope-binding domain in the cationic amino acid transporter determines the host range of ecotropic murine retroviruses. J Virol. 1993 Apr;67(4):2091–2096. doi: 10.1128/jvi.67.4.2091-2096.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andersen K. B. A domain of murine retrovirus surface protein gp70 mediates cell fusion, as shown in a novel SC-1 cell fusion system. J Virol. 1994 May;68(5):3175–3182. doi: 10.1128/jvi.68.5.3175-3182.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Arthos J., Deen K. C., Chaikin M. A., Fornwald J. A., Sathe G., Sattentau Q. J., Clapham P. R., Weiss R. A., McDougal J. S., Pietropaolo C. Identification of the residues in human CD4 critical for the binding of HIV. Cell. 1989 May 5;57(3):469–481. doi: 10.1016/0092-8674(89)90922-7. [DOI] [PubMed] [Google Scholar]
  4. Berg O. G., von Hippel P. H. Diffusion-controlled macromolecular interactions. Annu Rev Biophys Biophys Chem. 1985;14:131–160. doi: 10.1146/annurev.bb.14.060185.001023. [DOI] [PubMed] [Google Scholar]
  5. Bibb J. A., Witherell G., Bernhardt G., Wimmer E. Interaction of poliovirus with its cell surface binding site. Virology. 1994 May 15;201(1):107–115. doi: 10.1006/viro.1994.1270. [DOI] [PubMed] [Google Scholar]
  6. Camerini D., Seed B. A CD4 domain important for HIV-mediated syncytium formation lies outside the virus binding site. Cell. 1990 Mar 9;60(5):747–754. doi: 10.1016/0092-8674(90)90089-w. [DOI] [PubMed] [Google Scholar]
  7. Cao J., Bergeron L., Helseth E., Thali M., Repke H., Sodroski J. Effects of amino acid changes in the extracellular domain of the human immunodeficiency virus type 1 gp41 envelope glycoprotein. J Virol. 1993 May;67(5):2747–2755. doi: 10.1128/jvi.67.5.2747-2755.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Choppin J., Schaffar-Deshayes L., Debré P., Lévy J. P. Lymphoid cell surface receptor for Moloney leukemia virus envelope glycoprotein gp71. I. Binding characteristics. J Immunol. 1981 Jun;126(6):2347–2351. [PubMed] [Google Scholar]
  9. DeLarco J., Todaro G. J. Membrane receptors for murine leukemia viruses: characterization using the purified viral envelope glycoprotein, gp71. Cell. 1976 Jul;8(3):365–371. doi: 10.1016/0092-8674(76)90148-3. [DOI] [PubMed] [Google Scholar]
  10. Dimitrov D. S., Hillman K., Manischewitz J., Blumenthal R., Golding H. Correlation between kinetics of soluble CD4 interactions with HIV-1-Env-expressing cells and inhibition of syncytia formation: implications for mechanisms of cell fusion and therapy for AIDS. AIDS. 1992 Mar;6(3):249–256. doi: 10.1097/00002030-199203000-00001. [DOI] [PubMed] [Google Scholar]
  11. Dimitrov D. S., Hillman K., Manischewitz J., Blumenthal R., Golding H. Kinetics of soluble CD4 binding to cells expressing human immunodeficiency virus type 1 envelope glycoprotein. J Virol. 1992 Jan;66(1):132–138. doi: 10.1128/jvi.66.1.132-138.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Evans L. H., Morrison R. P., Malik F. G., Portis J., Britt W. J. A neutralizable epitope common to the envelope glycoproteins of ecotropic, polytropic, xenotropic, and amphotropic murine leukemia viruses. J Virol. 1990 Dec;64(12):6176–6183. doi: 10.1128/jvi.64.12.6176-6183.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ganguly K., Kalyanaraman V. S., Sarngadharan M. G. Analysis of the interaction between Rauscher murine leukemia virus and murine cell membrane receptor by in vitro binding assay. Cancer Lett. 1983 Feb;18(1):79–86. doi: 10.1016/0304-3835(83)90120-9. [DOI] [PubMed] [Google Scholar]
  14. Gelderblom H. R. Assembly and morphology of HIV: potential effect of structure on viral function. AIDS. 1991 Jun;5(6):617–637. [PubMed] [Google Scholar]
  15. Gray K. D., Roth M. J. Mutational analysis of the envelope gene of Moloney murine leukemia virus. J Virol. 1993 Jun;67(6):3489–3496. doi: 10.1128/jvi.67.6.3489-3496.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Heard J. M., Danos O. An amino-terminal fragment of the Friend murine leukemia virus envelope glycoprotein binds the ecotropic receptor. J Virol. 1991 Aug;65(8):4026–4032. doi: 10.1128/jvi.65.8.4026-4032.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kabat D. Cell surface receptors for ecotropic murine retroviruses: mobile membrane proteins that mediate binding and slow endocytosis of the viral envelope glycoprotein. Virology. 1989 Aug;171(2):467–474. doi: 10.1016/0042-6822(89)90616-8. [DOI] [PubMed] [Google Scholar]
  18. Kadan M. J., Sturm S., Anderson W. F., Eglitis M. A. Detection of receptor-specific murine leukemia virus binding to cells by immunofluorescence analysis. J Virol. 1992 Apr;66(4):2281–2287. doi: 10.1128/jvi.66.4.2281-2287.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kalyanaraman V. S., Sarngadharan M. G., Gallo R. C. Characterization of Rauscher murine leukemia virus envelope glycoprotein receptor in membranes from murine fibroblasts. J Virol. 1978 Dec;28(3):686–696. doi: 10.1128/jvi.28.3.686-696.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lasky L. A., Nakamura G., Smith D. H., Fennie C., Shimasaki C., Patzer E., Berman P., Gregory T., Capon D. J. Delineation of a region of the human immunodeficiency virus type 1 gp120 glycoprotein critical for interaction with the CD4 receptor. Cell. 1987 Sep 11;50(6):975–985. doi: 10.1016/0092-8674(87)90524-1. [DOI] [PubMed] [Google Scholar]
  21. Markowitz D., Goff S., Bank A. A safe packaging line for gene transfer: separating viral genes on two different plasmids. J Virol. 1988 Apr;62(4):1120–1124. doi: 10.1128/jvi.62.4.1120-1124.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Miller A. D., Rosman G. J. Improved retroviral vectors for gene transfer and expression. Biotechniques. 1989 Oct;7(9):980-2, 984-6, 989-90. [PMC free article] [PubMed] [Google Scholar]
  23. Moebius U., Clayton L. K., Abraham S., Harrison S. C., Reinherz E. L. The human immunodeficiency virus gp120 binding site on CD4: delineation by quantitative equilibrium and kinetic binding studies of mutants in conjunction with a high-resolution CD4 atomic structure. J Exp Med. 1992 Aug 1;176(2):507–517. doi: 10.1084/jem.176.2.507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Moore J. P., Klasse P. J. Thermodynamic and kinetic analysis of sCD4 binding to HIV-1 virions and of gp120 dissociation. AIDS Res Hum Retroviruses. 1992 Apr;8(4):443–450. doi: 10.1089/aid.1992.8.443. [DOI] [PubMed] [Google Scholar]
  25. Moore J. P., McKeating J. A., Norton W. A., Sattentau Q. J. Direct measurement of soluble CD4 binding to human immunodeficiency virus type 1 virions: gp120 dissociation and its implications for virus-cell binding and fusion reactions and their neutralization by soluble CD4. J Virol. 1991 Mar;65(3):1133–1140. doi: 10.1128/jvi.65.3.1133-1140.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Morgan R. A., Nussbaum O., Muenchau D. D., Shu L., Couture L., Anderson W. F. Analysis of the functional and host range-determining regions of the murine ectropic and amphotropic retrovirus envelope proteins. J Virol. 1993 Aug;67(8):4712–4721. doi: 10.1128/jvi.67.8.4712-4721.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ott D., Rein A. Basis for receptor specificity of nonecotropic murine leukemia virus surface glycoprotein gp70SU. J Virol. 1992 Aug;66(8):4632–4638. doi: 10.1128/jvi.66.8.4632-4638.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Shoup D., Szabo A. Role of diffusion in ligand binding to macromolecules and cell-bound receptors. Biophys J. 1982 Oct;40(1):33–39. doi: 10.1016/S0006-3495(82)84455-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Skov H., Andersen K. B. Mutational analysis of Moloney murine leukaemia virus surface protein gp70. J Gen Virol. 1993 Apr;74(Pt 4):707–714. doi: 10.1099/0022-1317-74-4-707. [DOI] [PubMed] [Google Scholar]
  30. Spies C. P., Compans R. W. Alternate pathways of secretion of simian immunodeficiency virus envelope glycoproteins. J Virol. 1993 Nov;67(11):6535–6541. doi: 10.1128/jvi.67.11.6535-6541.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Strand M., August J. T. Structural proteins of ribonucleic acid tumor viruses. Purification of envelope, core, and internal components. J Biol Chem. 1976 Jan 25;251(2):559–564. [PubMed] [Google Scholar]
  32. Wang H., Kavanaugh M. P., North R. A., Kabat D. Cell-surface receptor for ecotropic murine retroviruses is a basic amino-acid transporter. Nature. 1991 Aug 22;352(6337):729–731. doi: 10.1038/352729a0. [DOI] [PubMed] [Google Scholar]
  33. Wickham T. J., Shuler M. L., Hammer D. A., Granados R. R., Wood H. A. Equilibrium and kinetic analysis of Autographa californica nuclear polyhedrosis virus attachment to different insect cell lines. J Gen Virol. 1992 Dec;73(Pt 12):3185–3194. doi: 10.1099/0022-1317-73-12-3185. [DOI] [PubMed] [Google Scholar]
  34. Willey R. L., Smith D. H., Lasky L. A., Theodore T. S., Earl P. L., Moss B., Capon D. J., Martin M. A. In vitro mutagenesis identifies a region within the envelope gene of the human immunodeficiency virus that is critical for infectivity. J Virol. 1988 Jan;62(1):139–147. doi: 10.1128/jvi.62.1.139-147.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yoshimoto T., Yoshimoto E., Meruelo D. Identification of amino acid residues critical for infection with ecotropic murine leukemia retrovirus. J Virol. 1993 Mar;67(3):1310–1314. doi: 10.1128/jvi.67.3.1310-1314.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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