Structure of an inhibitor complex of the proteinase from feline immunodeficiency virus

Wlodawer, Alexander; Gustchina, Alla; Reshetnikova, L.; Łubkowski, J.; Zdanov, Alexander; Hui, Kwan Y.; Angleton, Eddie L.; Farmerie, William G.; Goodenow, Maureen M.; Bhatt, Deepa

doi:10.1038/nsb0695-480

Cited by 67 publications

(100 citation statements)

References 29 publications

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“…L97 80 is associated with the S1/S2 subsites, and substitutions at this position may influence the preferences at the P1/P2 positions of a substrate (20,28,54). In addition to poor/delayed cleavage at the FIV NC-p2 junction, inefficient processing at the FIV MA-CA cleavage junction by the L97 80 T mutant was also observed (Fig.…”

Section: Discussionmentioning

confidence: 89%

“…Most of the residues in the active sites of the two PRs are distinct (Fig. 1A, residues around Asp25 in HIV-1 PR and Asp 30 in FIV PR, the flaps, and the " 90s loop"), but the 3-dimensional structures of the two active cores are highly conserved (20,28,54). The structural locations of the substitutions investigated in this study are shown on one monomer of the dimeric FIV and HIV-1 PRs in Fig.…”

Section: Gag Proteins and Gag-pol Polyproteins Of The Two Viruses (Fimentioning

confidence: 99%

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Generation of Infectious Feline Immunodeficiency Virus (FIV) Encoding FIV/Human Immunodeficiency Virus Chimeric Protease

Lin¹,

Torbett

Elder³

2010

J Virol

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Feline immunodeficiency virus (FIV) and human immunodeficiency virus type 1 (HIV-1) proteases (PRs)share only 23% amino acid identity and exhibit distinct specificities yet have very similar 3-dimensional structures. Chimeric PRs in which HIV residues were substituted in structurally equivalent positions in FIV PR were prepared in order to study the molecular basis of PR specificity. Previous in vitro analyses showed that such substitutions dramatically altered the inhibitor specificity of mutant PRs but changed the rate and specificity of Gag cleavage so that chimeric FIVs were not infectious. Chimeric PRs encoding combinations of the I37V, N55M, M56I, V59I, L97T, I98P, Q99V, and P100N mutations were cloned into FIV Gag-Pol, and those constructs that best approximated the temporal cleavage pattern generated by wild-type FIV PR, while maintaining HIV-like inhibitor specificity, were selected. Two mutations, M56I and L97T, were intolerant to change and caused inefficient cleavage at NC-p2. However, a mutant PR with six substitutions (I37V, N55M, V59I, I98P, Q99V, and P100N) was selected and placed in the context of full-length FIV-34TF10. This virus, termed YCL6, had low-level infectivity ex vivo, and after passage, progeny that exhibited a higher growth rate emerged. The residue at the position of one of the six mutations, I98P, further mutated on passage to either P98H or P98S. Both PRs were sensitive to the HIV-1 PR inhibitors lopinavir (LPV) and darunavir (DRV), as well as to the broad-based inhibitor TL-3, with 50% inhibitory concentrations (IC 50 ) of 30 to 40 nM, consistent with ex vivo results obtained using mutant FIVs. The chimeras offer an infectivity system with which to screen compounds for potential as broad-based PR inhibitors, define structural parameters that dictate specificity, and investigate pathways for drug resistance development.

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Section: Discussionmentioning

confidence: 89%

Section: Gag Proteins and Gag-pol Polyproteins Of The Two Viruses (Fimentioning

confidence: 99%

Generation of Infectious Feline Immunodeficiency Virus (FIV) Encoding FIV/Human Immunodeficiency Virus Chimeric Protease

Lin¹,

Torbett

Elder³

2010

J Virol

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“…The structure of this inhibitor complexed with FIV proteinase has been solved and refined to 0.2-nm resolution [20]. Superposition of this complex with that of EIAV [Gly54]proteinase complexed with HBY-793 shows that the P3 naphthyalanine substituents occupy closely similar positions in the two structures.…”

Section: Discussionmentioning

confidence: 99%

Expression, Characterisation and Mutagenesis of the Aspartic Proteinase from Equine Infectious Anaemia Virus

Powell

Bur

Wlodawer

et al. 1996

European Journal of Biochemistry

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The gene encoding the proteinase from equine infectious anaemia virus (EIAV) was cloned and expressed in Escherichia coli. The recombinant EIAV proteinase was purified to homogeneity and shown to have the ability to process polyprotein and synthetic peptide substrates of human immunodeficiency virus (HIV) origin with an efficiency that can approach that exhibited by HIV proteinase. EIAV proteinase, however, was not susceptibfe to inhibition by a wide variety of inhibitors of HIV-1 proteinase, including those which have been licenced as anti-AIDS drugs. In this respect, EIAV proteinase behaves like an extreme case of a drug-resistant mutant of HIV-1 proteinase that has arisen under selective drug pressure. Only one potent inhibitor (HBY-793) of HIV-1 proteinase showed comparable efficiency against the EIAV enzyme; the compounds A-77003 and A-76889, which differ only in their stereochemistry and which are otherwise structurally identical to HBY-793 from residues P2 to P2' [nomenclature of Schechter, I.

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“…It was clear that in EIAV PR the variable loops A1 and A2 would be the same as in the HIV-1 enzyme. We expected the insertion of five residues in EIAV PR to be located in the variable region of a wide surface loop, which had been observed to have a very specific conformation in all known structures of the retroviral proteinases Tong et al, 1993;Wlodawer et al, 1995). Because we expected this fragment to have a different conformation in EIAV PR as well, we decided not to model it, and instead kept it as polyalanine with the conformation found in HIV-1 PR, without inserting extra residues.…”

Section: Model Buildingmentioning

confidence: 99%

“…and for the proteinases of other retroviruses [e.g., SIV, Sugrue et al, 1994; feline immunodeficiency virus (FIV), Wlodawer et al, 1995; Rous sarcoma virus (RSV), Jaskolski et al, 19901. and discussion We report here the elucidation of the structure of a single-site I54G mutant form of EIAV PR complexed with the only inhibStructure solution and de.ycrjptjon itor (HBY-793) known to inhibit both the wild-type and mutant EIAV PR, as well as HIV-1 PR, with subnanomolar potency…”

mentioning

confidence: 99%

Structure of equine infectious anemia virus proteinase complexed with an inhibitor

et al. 1996

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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 I54G 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 a-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.

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Structure of an inhibitor complex of the proteinase from feline immunodeficiency virus

Cited by 67 publications

References 29 publications

Generation of Infectious Feline Immunodeficiency Virus (FIV) Encoding FIV/Human Immunodeficiency Virus Chimeric Protease

Generation of Infectious Feline Immunodeficiency Virus (FIV) Encoding FIV/Human Immunodeficiency Virus Chimeric Protease

Expression, Characterisation and Mutagenesis of the Aspartic Proteinase from Equine Infectious Anaemia Virus

Structure of equine infectious anemia virus proteinase complexed with an inhibitor

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