Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Cellular receptor structures for pseudorabies virus are blocked by antithrombin III

Abstract

Pseudorabies virus (PrV), an alphaherpesvirus of swine, uses cellular heparan sulfate residues as a receptor for attachment. Interaction of the virus with its receptor is mediated by the envelope glycoprotein C (PrV-gC), a protein with heparin-binding properties. We have previously shown that a region of this protein shows structural similarities to the high-affinity heparin-binding site of the serum protease-inhibitor antithrombin III (ATII). In this publication, we describe the effect of ATIII on interaction of PrV with its cellular receptor. ATIII bound specifically to heparan sulfate residues on the surface of herpesvirus-permissive RK13 cells. Binding of ATIII to RK13 cells interfered with adsorption of radioactively labelled PrV to these cells. Enzymatic treatment using heparinase I (E.C. 4.2.2.7) removed the receptor for PrV as well as the receptor for ATIII. Since amino acids 130–137 of the high-affinity heparin-binding site of ATIII show structural similarities to amino acids 134–141 of PrV-gC, both sequences were synthesized as synthetic peptides. Although interaction of the peptide derived from ATIII with heparin was significantly stronger, both peptides interacted specifically with heparin in assays in vitro. These results suggest that PrV and ATIII interact with the same structure on the cellular surface.

This is a preview of subscription content, log in to check access.

References

  1. 1.

    Agostini AI de, Watkins SC, Slayter HS, Youssoufian H, Rosenberg RD (1990) Localization of anticoagulantly active heparan sulfate proteoglycans in vascular endothelium: antithrombin binding on cultured endothelial cells and perfused rat aorta. J Cell Biol 111:1293–1304

  2. 2.

    Cardin AD, Weintraub HJR (1989) Molecular modeling of protein-glycosaminoglycan interactions. Arteriosclerosis 9:21–32

  3. 3.

    Evans DLI, Marshall CJ, Christey PB, Carrell RW (1992) Heparin binding site, conformational change, and activation of antithrombin. Biochem 31:12629–12642

  4. 4.

    Flynn SJ, Ryan P (1995) A heterologous heparin-binding domain can promote functional attachment of a pseudorabies virus gC mutant to cell surfaces. J Virol 69:834–839

  5. 5.

    Fuller AO, Spear PG (1985) Specifities of monoclonal and polyclonal antibodies that inhibit adsorption of herpes simplex virus to cells and lack of inhibition by potent neutralizing antibodies. J Virol 55:475–482

  6. 6.

    Geysen HM, Barteling SJ, Meloen RH (1985) Small peptides induces antibodies with a sequence and structural requirement for binding antigen comparable to antibodies raised against the native protein. Proc Natl Acad Sci USA 82:178–182

  7. 7.

    Griffith MJ (1982) Kinetics of the heparin-enhanced antithrombin II/thrombin reaction. Evidence for a template model for the mechanism of the action of heparin. J Biol Chem 257: 7360–7365

  8. 8.

    Hampl H, Ben-Porat T, Ehrlicher L, Habermehl K-O, Kaplan AS (1984) Characterization of the envelope proteins of pseudorabies virus. J Virol 52:583–590

  9. 9.

    Herold BC, WuDunn D, Soltys N, Spear PG (1991) Glycoprotein C of herpes simplex virus type 1 plays a principal role in the adsorption of virus to cells and in infectivity. J Virol 65:1090–1098

  10. 10.

    Hirose N, Krivanek M, Jackson RL, Cardin AD (1986) A dotblot assay for heparin-binding proteins. Anal Biochem 156:320–325

  11. 11.

    Jackson RL, Busch SJ, Cardin AD (1991) Glycosaminoglycans: molecular properties, proteins interactions, and role in physiological processes. Physiol Rev 71:481–539

  12. 12.

    Karger A, Mettenleiter TC (1993) Glycoproteins gIII and gp50 play dominant roles in the biphasic attachment of pseudorabies virus. Virology 194:654–664

  13. 13.

    Kuhn LA, Griffin JH, Fisher CL, Greengard JS, Bourma BN, Espana F, Tainer JA (1990) Elucidating the structural chemistry of glycosaminoglycan recognition by protein C inhibitor. Proc Natl Acad Sci USA 87:8506–8510

  14. 14.

    Marsh M, Helenius A (198) Virus entry into animal cells. Adv Virus Res 36:107–151

  15. 15.

    Mettenleiter TC, Zsak L, Zuckermann F, Sugg N, Kern H, Benporat T (1990) Interaction of glycoprotein gIII with a cellular heparin-like substance mediates adsorption of pseudorabies virus. J Virol 64:278–286

  16. 16.

    Mourey L, Samama J-P, Delarue M, Peptitiou M, Choay J, Moras D (1993) Crystal structure of cleaved bovine antithrombin III at 3.2 Å resolution. J Mol Biol 232:223–241

  17. 17.

    Sawitzky D, Hampl H, Habermehl K-O (1990) Comparison of heparin-sensitive attachment of pseudorabies virus and herpes simplex virus type 1 and identification of heparin-binding PrV-glycoproteins. J Gen Virol 71:1221–1225

  18. 18.

    Sawitzky D, Hampl H, Habermehl K-O (1990) Entry of pseudorabies virus into CHO cells is blocked at the level of penetration. Arch Virol 115:309–316

  19. 19.

    Sawitzky D, Voigt A, Habermehl K-O (1993) A peptide-model for the heparin-binding property of pseudorabies virus glycoprotein III. Med Microbiol Immunol 182:1273–1281

  20. 20.

    Shieh M-T, WuDunn D, Montgomery RI, Esko JD, Spear PG (1992) Cell surface receptors for herpes simplex virus are heparan sulfate proteoglycans. J Cell Biol 116:1273–1281

  21. 21.

    Silva ME, Dietrich CP (1975) Structure of heparin. Characterisation of the products from heparin by the action of a heparinase and a heparitinase from Flavobacterium heparinum. J Biol Chem 250:6841–6846

  22. 22.

    Tyler-Cross R, Sobel M, Marques D, Harris RB (1994) Heparin binding domain peptides of antithrombin III: analysis by isothermal titration calorimetry and circular dichroism spectroscopy. Protein Sci 3:620–627

  23. 23.

    WuDunn D, Spear PG (1989) Initial interaction of herpes simplex virus with cells is binding to heparan sulfate. J Virol 63:52–58

  24. 24.

    Zuckermann F, Zsak L, Reilly L, Sugg N, Ben-Porat T (1989) Early interactions of pseudorabies virus with host cell: function of glycoprotein gIII. J Virol 63:3323–3329

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Voigt, A., Sawitzky, D., Zeichhardt, H. et al. Cellular receptor structures for pseudorabies virus are blocked by antithrombin III. Med Microbiol Immunol 184, 97–103 (1995). https://doi.org/10.1007/BF00221393

Download citation

Keywords

  • Peptide
  • Sulfate
  • Heparin
  • Synthetic Peptide
  • Antithrombin