FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Protein Sci / Structure of equine infectious anemia virus proteinase compl...
Protein science : a publication of the Protein Society1996; 5(8); 1453-1465; doi: 10.1002/pro.5560050802

Structure of equine infectious anemia virus proteinase complexed with an inhibitor.

Abstract: Equine infectious anemia virus (EIAV), the causative agent of infectious anemia in horses, is a member of the lentiviral family. The virus-encoded proteinase (PR) processes viral polyproteins into functional molecules during replication and it also cleaves viral nucleocapsid protein during infection. The X-ray structure of a complex of the 154G mutant of EIAV PR with the inhibitor HBY-793 was solved at 1.8 A resolution and refined to a crystallographic R-factor of 0.136. The molecule is a dimer in which the monomers are related by a crystallographic twofold axis. Although both the enzyme and the inhibitor are symmetric, the interactions between the central part of the inhibitor and the active site aspartates are asymmetric, and the inhibitor and the two flaps are partially disordered. The overall fold of EIAV PR is very similar to that of other retroviral proteinases. However, a novel feature of the EIAV PR structure is the appearance of the second alpha-helix in the monomer in a position predicted by the structural template for the family of aspartic proteinases. The parts of the EIAV PR with the highest resemblance to human immunodeficiency virus type 1 PR include the substrate-binding sites; thus, the differences in the specificity of both enzymes have to be explained by enzyme-ligand interactions at the periphery of the active site as well.
Get Full Text
Publication Date: 1996-08-01 PubMed ID: 8844837PubMed Central: PMC2143478DOI: 10.1002/pro.5560050802Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Comparative Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research study presents the experimental exploration of the structure of a proteinase from the equine infectious anemia virus (EIAV), and how it interplays with a specific inhibitor. The authors utilized X-ray crystallography to decipher this structure at an atomic level, which offered insights into the similarities between this proteinase and others in retroviruses.

Study Objective and Methodology

  • The main objective of this study was to understand the structure of the equine infectious anemia virus (EIAV) proteinase (PR) and its interaction with an inhibitor (HBY-793). The EIAV PR plays a significant role in viral replication by processing viral polyproteins into functional molecules and cleaving viral nucleocapsid protein.
  • The researchers utilized the tool of X-ray crystallography to achieve this. They specifically examined the complex of the 154G mutant of EIAV PR with the inhibitor at 1.8 A resolution.

Findings and Analysis

  • Upon analysis, the researchers found that EIAV PR is a dimer, a complex of two identical molecules (monomers). The monomers are related by a twofold axis, maintaining a symmetrical structure.
  • Interestingly, despite the symmetric nature of the enzyme and the inhibitor, the interactions between the central part of the inhibitor and the active site aspartates were found to be asymmetrical. Additionally, the inhibitor and the two flaps were partially disordered.
  • The EIAV PR structure shared striking similarities with other retroviral proteinases, but it also had a unique feature: the appearance of the second alpha-helix in the monomer in a position predicted by the structural template for the family of aspartic proteinases.
  • The areas of the EIAV PR most resembling human immunodeficiency virus type 1 PR were the substrate-binding sites. Moreover, the researchers found that differences in the specificity of both enzymes had to be linked also to enzyme-ligand interactions at the periphery of the active site.

Implications of the Study

  • This study deepens our understanding of the structure and functioning of EIAV PR and how inhibitors work against it. This knowledge can play a pivotal role in developing more effective antiviral drugs.
  • By elucidating the similarities and differences between EIAV PR and the similar proteinase in HIV, this research could shed light on new potential lines of approach in HIV/AIDS treatment and prevention.

Cite This Article

APA
Gustchina A, Kervinen J, Powell DJ, Zdanov A, Kay J, Wlodawer A. (1996). Structure of equine infectious anemia virus proteinase complexed with an inhibitor. Protein Sci, 5(8), 1453-1465. https://doi.org/10.1002/pro.5560050802

Publication

Protein science : a publication of the Protein Society
ISSN: 0961-8368
NlmUniqueID: 9211750
Country: United States
Language: English
Volume: 5
Issue: 8
Pages: 1453-1465

Researcher Affiliations

Gustchina, A
  • Macromolecular Structure Laboratory, NCI-Frederick Cancer Research and Development Center, Maryland 21702, USA.
Kervinen, J
    Powell, D J
      Zdanov, A
        Kay, J
          Wlodawer, A

            MeSH Terms

            • Amino Acid Sequence
            • Animals
            • Binding Sites
            • Crystallography, X-Ray
            • Endopeptidases / chemistry
            • Endopeptidases / genetics
            • Escherichia coli / genetics
            • HIV Protease Inhibitors / chemistry
            • HIV-1 / chemistry
            • Horses
            • Humans
            • Hydrogen Bonding
            • Infectious Anemia Virus, Equine / enzymology
            • Infectious Anemia Virus, Equine / genetics
            • Molecular Sequence Data
            • Protease Inhibitors / chemistry
            • Protein Structure, Secondary
            • Protein Structure, Tertiary
            • Pyridines / chemistry
            • Recombinant Proteins / chemistry
            • Recombinant Proteins / genetics
            • Recombinant Proteins / isolation & purification
            • Retroviridae Proteins / antagonists & inhibitors
            • Retroviridae Proteins / chemistry
            • Retroviridae Proteins / genetics
            • Sequence Alignment

            References

            This article includes 31 references
            1. Andreeva NS, Gustchina AE. On the supersecondary structure of acid proteases.. Biochem Biophys Res Commun 1979 Mar 15;87(1):32-42.
              pubmed: 36889doi: 10.1016/0006-291x(79)91643-7google scholar: lookup
            2. Cheevers WP, McGuire TC. Equine infectious anemia virus: immunopathogenesis and persistence.. Rev Infect Dis 1985 Jan-Feb;7(1):83-8.
              pubmed: 2984759doi: 10.1093/clinids/7.1.83google scholar: lookup
            3. Jones TA. Diffraction methods for biological macromolecules. Interactive computer graphics: FRODO.. Methods Enzymol 1985;115:157-71.
              pubmed: 3841179doi: 10.1016/0076-6879(85)15014-7google scholar: lookup
            4. Hendrickson WA. Stereochemically restrained refinement of macromolecular structures.. Methods Enzymol 1985;115:252-70.
              pubmed: 3841182doi: 10.1016/0076-6879(85)15021-4google scholar: lookup
            5. Stephens RM, Casey JW, Rice NR. Equine infectious anemia virus gag and pol genes: relatedness to visna and AIDS virus.. Science 1986 Feb 7;231(4738):589-94.
              pubmed: 3003905doi: 10.1126/science.3003905google scholar: lookup
            6. Satow Y, Cohen GH, Padlan EA, Davies DR. Phosphocholine binding immunoglobulin Fab McPC603. An X-ray diffraction study at 2.7 A.. J Mol Biol 1986 Aug 20;190(4):593-604.
              pubmed: 3097327doi: 10.1016/0022-2836(86)90245-7google scholar: lookup
            7. Wlodawer A, Miller M, Jaskólski M, Sathyanarayana BK, Baldwin E, Weber IT, Selk LM, Clawson L, Schneider J, Kent SB. Conserved folding in retroviral proteases: crystal structure of a synthetic HIV-1 protease.. Science 1989 Aug 11;245(4918):616-21.
              pubmed: 2548279doi: 10.1126/science.2548279google scholar: lookup
            8. Rao JK, Wlodawer A. Is the pseudo-dyad in retroviral proteinase monomers structural or evolutionary?. FEBS Lett 1990 Jan 29;260(2):201-5.
              pubmed: 2153583doi: 10.1016/0014-5793(90)80103-pgoogle scholar: lookup
            9. Jaskólski M, Miller M, Rao JK, Leis J, Wlodawer A. Structure of the aspartic protease from Rous sarcoma retrovirus refined at 2-A resolution.. Biochemistry 1990 Jun 26;29(25):5889-98.
              pubmed: 2166563doi: 10.1021/bi00477a002google scholar: lookup
            10. Tözsér J, Gustchina A, Weber IT, Blaha I, Wondrak EM, Oroszlan S. Studies on the role of the S4 substrate binding site of HIV proteinases.. FEBS Lett 1991 Feb 25;279(2):356-60.
              pubmed: 2001747doi: 10.1016/0014-5793(91)80186-7google scholar: lookup
            11. Roberts MM, Copeland TD, Oroszlan S. In situ processing of a retroviral nucleocapsid protein by the viral proteinase.. Protein Eng 1991 Aug;4(6):695-700.
              pubmed: 1658777doi: 10.1093/protein/4.6.695google scholar: lookup
            12. Gustchina A, Weber IT. Comparative analysis of the sequences and structures of HIV-1 and HIV-2 proteases.. Proteins 1991;10(4):325-39.
              pubmed: 1946342doi: 10.1002/prot.340100406google scholar: lookup
            13. Andreeva NS. A consensus template for the aspartic proteinase fold.. Adv Exp Med Biol 1991;306:559-72.
              pubmed: 1812759doi: 10.1007/978-1-4684-6012-4_77google scholar: lookup
            14. Griffiths JT, Phylip LH, Konvalinka J, Strop P, Gustchina A, Wlodawer A, Davenport RJ, Briggs R, Dunn BM, Kay J. Different requirements for productive interaction between the active site of HIV-1 proteinase and substrates containing -hydrophobic*hydrophobic- or -aromatic*pro- cleavage sites.. Biochemistry 1992 Jun 9;31(22):5193-200.
              pubmed: 1606143doi: 10.1021/bi00137a015google scholar: lookup
            15. Tözsér J, Friedman D, Weber IT, Bláha I, Oroszlan S. Studies on the substrate specificity of the proteinase of equine infectious anemia virus using oligopeptide substrates.. Biochemistry 1993 Apr 6;32(13):3347-53.
              pubmed: 8384879doi: 10.1021/bi00064a018google scholar: lookup
            16. Weber IT, Tözsér J, Wu J, Friedman D, Oroszlan S. Molecular model of equine infectious anemia virus proteinase and kinetic measurements for peptide substrates with single amino acid substitutions.. Biochemistry 1993 Apr 6;32(13):3354-62.
              pubmed: 8384880doi: 10.1021/bi00064a019google scholar: lookup
            17. Thanki N, Rao JK, Foundling SI, Howe WJ, Moon JB, Hui JO, Tomasselli AG, Heinrikson RL, Thaisrivongs S, Wlodawer A. Crystal structure of a complex of HIV-1 protease with a dihydroxyethylene-containing inhibitor: comparisons with molecular modeling.. Protein Sci 1992 Aug;1(8):1061-72.
              pubmed: 1304383doi: 10.1002/pro.5560010811google scholar: lookup
            18. Wlodawer A, Erickson JW. Structure-based inhibitors of HIV-1 protease.. Annu Rev Biochem 1993;62:543-85.
              pubmed: 8352596doi: 10.1146/annurev.bi.62.070193.002551google scholar: lookup
            19. Otto MJ, Garber S, Winslow DL, Reid CD, Aldrich P, Jadhav PK, Patterson CE, Hodge CN, Cheng YS. In vitro isolation and identification of human immunodeficiency virus (HIV) variants with reduced sensitivity to C-2 symmetrical inhibitors of HIV type 1 protease.. Proc Natl Acad Sci U S A 1993 Aug 15;90(16):7543-7.
              pubmed: 8356053doi: 10.1073/pnas.90.16.7543google scholar: lookup
            20. Tong L, Pav S, Pargellis C, Dô F, Lamarre D, Anderson PC. Crystal structure of human immunodeficiency virus (HIV) type 2 protease in complex with a reduced amide inhibitor and comparison with HIV-1 protease structures.. Proc Natl Acad Sci U S A 1993 Sep 15;90(18):8387-91.
              pubmed: 8378311doi: 10.1073/pnas.90.18.8387google scholar: lookup
            21. Griffiths JT, Tomchak LA, Mills JS, Graves MC, Cook ND, Dunn BM, Kay J. Interactions of substrates and inhibitors with a family of tethered HIV-1 and HIV-2 homo- and heterodimeric proteinases.. J Biol Chem 1994 Feb 18;269(7):4787-93.
              pubmed: 8106448
            22. Sugrue RJ, Almond N, Kitchin P, Richardson SM, Wilderspin AF. Purification of crystallizable recombinant SIVmac251-32H proteinase.. Protein Expr Purif 1994 Feb;5(1):76-83.
              pubmed: 8167477doi: 10.1006/prep.1994.1011google scholar: lookup
            23. Gustchina A, Sansom C, Prevost M, Richelle J, Wodak SY, Wlodawer A, Weber IT. Energy calculations and analysis of HIV-1 protease-inhibitor crystal structures.. Protein Eng 1994 Mar;7(3):309-17.
              pubmed: 8177879doi: 10.1093/protein/7.3.309google scholar: lookup
            24. Daenke S, Schramm HJ, Bangham CR. Analysis of substrate cleavage by recombinant protease of human T cell leukaemia virus type 1 reveals preferences and specificity of binding.. J Gen Virol 1994 Sep;75 ( Pt 9):2233-9.
              pubmed: 8077922doi: 10.1099/0022-1317-75-9-2233google scholar: lookup
            25. Markowitz M, Mo H, Kempf DJ, Norbeck DW, Bhat TN, Erickson JW, Ho DD. Selection and analysis of human immunodeficiency virus type 1 variants with increased resistance to ABT-538, a novel protease inhibitor.. J Virol 1995 Feb;69(2):701-6.
              pubmed: 7815532doi: 10.1128/JVI.69.2.701-706.1995google scholar: lookup
            26. Condra JH, Schleif WA, Blahy OM, Gabryelski LJ, Graham DJ, Quintero JC, Rhodes A, Robbins HL, Roth E, Shivaprakash M. In vivo emergence of HIV-1 variants resistant to multiple protease inhibitors.. Nature 1995 Apr 6;374(6522):569-71.
              pubmed: 7700387doi: 10.1038/374569a0google scholar: lookup
            27. Baldwin ET, Bhat TN, Liu B, Pattabiraman N, Erickson JW. Structural basis of drug resistance for the V82A mutant of HIV-1 proteinase.. Nat Struct Biol 1995 Mar;2(3):244-9.
              pubmed: 7773792doi: 10.1038/nsb0395-244google scholar: lookup
            28. Nicholson LK, Yamazaki T, Torchia DA, Grzesiek S, Bax A, Stahl SJ, Kaufman JD, Wingfield PT, Lam PY, Jadhav PK. Flexibility and function in HIV-1 protease.. Nat Struct Biol 1995 Apr;2(4):274-80.
              pubmed: 7796263doi: 10.1038/nsb0495-274google scholar: lookup
            29. Gulnik SV, Suvorov LI, Liu B, Yu B, Anderson B, Mitsuya H, Erickson JW. Kinetic characterization and cross-resistance patterns of HIV-1 protease mutants selected under drug pressure.. Biochemistry 1995 Jul 25;34(29):9282-7.
              pubmed: 7626598doi: 10.1021/bi00029a002google scholar: lookup
            30. Wlodawer A, Gustchina A, Reshetnikova L, Lubkowski J, Zdanov A, Hui KY, Angleton EL, Farmerie WG, Goodenow MM, Bhatt D. Structure of an inhibitor complex of the proteinase from feline immunodeficiency virus.. Nat Struct Biol 1995 Jun;2(6):480-8.
              pubmed: 7664111doi: 10.1038/nsb0695-480google scholar: lookup
            31. Erickson JW. The not-so-great escape.. Nat Struct Biol 1995 Jul;2(7):523-9.
              pubmed: 7664118doi: 10.1038/nsb0795-523google scholar: lookup

            Citations

            This article has been cited 15 times.
            1. Golda M, Mótyán JA, Nagy K, Matúz K, Nagy T, Tőzsér J. Biochemical Characterization of Human Retroviral-Like Aspartic Protease 1 (ASPRV1).. Biomolecules 2020 Jul 6;10(7).
              doi: 10.3390/biom10071004pubmed: 32640672google scholar: lookup
            2. Golda M, Mótyán JA, Mahdi M, Tőzsér J. Functional Study of the Retrotransposon-Derived Human PEG10 Protease.. Int J Mol Sci 2020 Mar 31;21(7).
              doi: 10.3390/ijms21072424pubmed: 32244497google scholar: lookup
            3. Mótyán JA, Miczi M, Tőzsér J. Dimer Interface Organization is a Main Determinant of Intermonomeric Interactions and Correlates with Evolutionary Relationships of Retroviral and Retroviral-Like Ddi1 and Ddi2 Proteases.. Int J Mol Sci 2020 Feb 17;21(4).
              doi: 10.3390/ijms21041352pubmed: 32079302google scholar: lookup
            4. Gazda LD, Joóné Matúz K, Nagy T, Mótyán JA, Tőzsér J. Biochemical characterization of Ty1 retrotransposon protease.. PLoS One 2020;15(1):e0227062.
              doi: 10.1371/journal.pone.0227062pubmed: 31917798google scholar: lookup
            5. Li M, Gustchina A, Cruz R, Simões M, Curto P, Martinez J, Faro C, Simões I, Wlodawer A. Structure of RC1339/APRc from Rickettsia conorii, a retropepsin-like aspartic protease.. Acta Crystallogr D Biol Crystallogr 2015 Oct;71(Pt 10):2109-18.
              doi: 10.1107/S1399004715013905pubmed: 26457434google scholar: lookup
            6. Jaskolski M, Miller M, Mohana Rao JK, Gustchina A, Wlodawer A. Elucidation of the structure of retroviral proteases: a reminiscence.. FEBS J 2015 Nov;282(21):4059-66.
              doi: 10.1111/febs.13397pubmed: 26258480google scholar: lookup
            7. Zhang S, Kaplan AH, Tropsha A. HIV-1 protease function and structure studies with the simplicial neighborhood analysis of protein packing method.. Proteins 2008 Nov 15;73(3):742-53.
              doi: 10.1002/prot.22094pubmed: 18498108google scholar: lookup
            8. Li M, Laco GS, Jaskolski M, Rozycki J, Alexandratos J, Wlodawer A, Gustchina A. Crystal structure of human T cell leukemia virus protease, a novel target for anticancer drug design.. Proc Natl Acad Sci U S A 2005 Dec 20;102(51):18332-7.
              doi: 10.1073/pnas.0509335102pubmed: 16352712google scholar: lookup
            9. Bagossi P, Sperka T, Fehér A, Kádas J, Zahuczky G, Miklóssy G, Boross P, Tözsér J. Amino acid preferences for a critical substrate binding subsite of retroviral proteases in type 1 cleavage sites.. J Virol 2005 Apr;79(7):4213-8.
              doi: 10.1128/JVI.79.7.4213-4218.2005pubmed: 15767422google scholar: lookup
            10. Davis DA, Brown CA, Newcomb FM, Boja ES, Fales HM, Kaufman J, Stahl SJ, Wingfield P, Yarchoan R. Reversible oxidative modification as a mechanism for regulating retroviral protease dimerization and activation.. J Virol 2003 Mar;77(5):3319-25.
              doi: 10.1128/jvi.77.5.3319-3325.2003pubmed: 12584357google scholar: lookup
            11. Dunn BM, Goodenow MM, Gustchina A, Wlodawer A. Retroviral proteases.. Genome Biol 2002;3(4):REVIEWS3006.
              doi: 10.1186/gb-2002-3-4-reviews3006pubmed: 11983066google scholar: lookup
            12. Pfrepper KI, Reed J, Rackwitz HR, Schnölzer M, Flügel RM. Characterization of peptide substrates and viral enzyme that affect the cleavage site specificity of the human spumaretrovirus proteinase.. Virus Genes 2001 Jan;22(1):61-72.
              doi: 10.1023/a:1008134419542pubmed: 11210941google scholar: lookup
            13. Tözsér J. Specificity of Retroviral Proteinases Based on Substrates Containing Tyrosine and Proline at the Site of Cleavage.. Pathol Oncol Res 1997;3(2):142-146.
              doi: 10.1007/BF02907811pubmed: 11173643google scholar: lookup
            14. Kervinen J, Lubkowski J, Zdanov A, Bhatt D, Dunn BM, Hui KY, Powell DJ, Kay J, Wlodawer A, Gustchina A. Toward a universal inhibitor of retroviral proteases: comparative analysis of the interactions of LP-130 complexed with proteases from HIV-1, FIV, and EIAV.. Protein Sci 1998 Nov;7(11):2314-23.
              doi: 10.1002/pro.5560071108pubmed: 9827997google scholar: lookup
            15. Laco GS, Fitzgerald MC, Morris GM, Olson AJ, Kent SB, Elder JH. Molecular analysis of the feline immunodeficiency virus protease: generation of a novel form of the protease by autoproteolysis and construction of cleavage-resistant proteases.. J Virol 1997 Jul;71(7):5505-11.
              doi: 10.1128/JVI.71.7.5505-5511.1997pubmed: 9188624google scholar: lookup
            Subscribe to our newsletter
            Join 99,357 horse owners like you who receive our email updates on horse health and nutrition.