Investigation of an Allosteric Site of HIV-1 Proteinase Involved in Inhibition by Cu2+

Danielson, Helena; Lindgren, Maria T.; Markgren, Per-Olof; Nillroth, Ulrika

doi:10.1007/978-1-4615-5373-1_13

Cited by 23 publications

(19 citation statements)

References 11 publications

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“…Wild-type HIV-1 protease (HXB2 clone) and its three mutants, the I84V, L90M, and I84V þ L90M genes, were a kind gift of Professor Helena Danielson, Uppsala University. Protease expression, isolation, refolding, and analysis were performed as previously described [72].…”

Section: Methodsmentioning

confidence: 99%

A Look inside HIV Resistance through Retroviral Protease Interaction Maps

Kontijevskis¹,

Prūsis²,

Petrovska³

et al. 2005

PLoS Comput Biol

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Retroviruses affect a large number of species, from fish and birds to mammals and humans, with global socioeconomic negative impacts. Here the authors report and experimentally validate a novel approach for the analysis of the molecular networks that are involved in the recognition of substrates by retroviral proteases. Using multivariate analysis of the sequence-based physiochemical descriptions of 61 retroviral proteases comprising wild-type proteases, natural mutants, and drug-resistant forms of proteases from nine different viral species in relation to their ability to cleave 299 substrates, the authors mapped the physicochemical properties and cross-dependencies of the amino acids of the proteases and their substrates, which revealed a complex molecular interaction network of substrate recognition and cleavage. The approach allowed a detailed analysis of the molecular-chemical mechanisms involved in substrate cleavage by retroviral proteases.Citation: Kontijevskis A, Prusis P, Petrovska R, Yahorava S, Mutulis F, et al. (2007) A look inside HIV resistance through retroviral protease interaction maps. PLoS Comput Biol 3(3): e48.

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Section: Methodsmentioning

confidence: 99%

A Look inside HIV Resistance through Retroviral Protease Interaction Maps

Kontijevskis¹,

Prūsis²,

Petrovska³

et al. 2005

PLoS Comput Biol

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“…DRV has proved highly effective in salvage therapy for patients failing other treatments 5. Success of the design strategy was verified by crystallographic and kinetic analysis of DRV with HIV-1 protease and its mutants 6,7,8,9,10…”

mentioning

confidence: 98%

Solution Kinetics Measurements Suggest HIV-1 Protease Has Two Binding Sites for Darunavir and Amprenavir

Kovalevsky

Ghosh²,

Weber³

2008

J. Med. Chem.

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Darunavir, a potent antiviral drug, showed an unusual second binding site on the HIV-1 protease dimer surface of the V32I drug resistant mutant and normal binding in the active site cavity. Kinetic analysis for wild type and mutant protease showed mixed-type competitive-uncompetitive inhibition for darunavir and the chemically related amprenavir, while saquinavir showed competitive inhibition. The inhibition model is consistent with the observed second binding site for darunavir and helps to explain its antiviral potency.HIV-1 protease (PR) has been the target of intensive research for the past two decades that has led to effective drugs for the treatment of HIV/AIDS, i.e. small molecule synthetic protease inhibitors (PIs). 1 The PIs have become the paradigm for successful structure-assisted drug design, 2 with nine PIs now approved by the FDA for AIDS therapy. The addition of PIs to the antiviral regimens resulted in highly increased survival rates and lower morbidity caused by the disease in the past decade. However, the emergence of drug-resistant HIV has necessitated the recent development of PIs, such as darunavir (DRV) designed to target PR mutants. 3,4 DRV has proved highly effective in salvage therapy for patients failing other treatments. 5 Success of the design strategy was verified by crystallographic and kinetic analysis of DRV with HIV-1 protease and its mutants. 6,7,8,9,10 The clinical PIs have been designed to inhibit the activity of the HIV-1 PR by competitively binding inside the active-site cavity, 11,12,13 which is formed by a dimer of two identical 99-residue subunits. Numerous crystal structures of HIV-1 PR complexed with different PIs have shown exclusive binding of the inhibitors in the active-site cavity. Some reversible inhibitor molecules, like beta-lactam compounds 14 and Nb-containing polyoxometalates 15 , were proposed to bind on the surface of the enzyme in uncompetitive or non-competitive manner, respectively, rather than in the usual active site cavity. Also, irreversible inhibitors like haloperidol analogues 16 and Cu 2+ ions 7 can disrupt the HIV-1 PR activity by attaching covalently at surface sites. One peptide inhibitor was observed to bind on the protein surface between PR dimers in the crystal lattice, although this inhibitor showed no specific interactions with PR. 17 An unusual second binding site for DRV was observed in the two high-resolution crystal structures of complexes with HIV-1 PR mutants with the single substitutions of V32I (0.84 Å) # to whom correspondence should be addressed. and M46L (1.22 Å). 18 In both structures one inhibitor molecule occupied the active-site cavity, while the other inhibitor molecule was located in a deep groove on the PR surface in the flap region. Thus the PR/DRV 2 species was observed in the solid state ( Figure 1a). The surfacebound inhibitor molecule formed a number of specific interactions with the mutant PR residues, including hydrogen bonds (Figure 1b), C-H…O and C-H…π contacts, unlike the peptide observed on the flap sur...

show abstract

“…Protease expression, isolation, refolding, and analysis were performed as previously described [72]. …”

Section: Methodsmentioning

confidence: 99%

A Look Inside HIV Resistance through Retroviral Protease Interaction Maps

et al. 2007

View full text Add to dashboard Cite

Retroviruses affect a large number of species, from fish and birds to mammals and humans, with global socioeconomic negative impacts. Here the authors report and experimentally validate a novel approach for the analysis of the molecular networks that are involved in the recognition of substrates by retroviral proteases. Using multivariate analysis of the sequence-based physiochemical descriptions of 61 retroviral proteases comprising wild-type proteases, natural mutants, and drug-resistant forms of proteases from nine different viral species in relation to their ability to cleave 299 substrates, the authors mapped the physicochemical properties and cross-dependencies of the amino acids of the proteases and their substrates, which revealed a complex molecular interaction network of substrate recognition and cleavage. The approach allowed a detailed analysis of the molecular–chemical mechanisms involved in substrate cleavage by retroviral proteases.

show abstract

Investigation of an Allosteric Site of HIV-1 Proteinase Involved in Inhibition by Cu2+

Cited by 23 publications

References 11 publications

A Look inside HIV Resistance through Retroviral Protease Interaction Maps

A Look inside HIV Resistance through Retroviral Protease Interaction Maps

Solution Kinetics Measurements Suggest HIV-1 Protease Has Two Binding Sites for Darunavir and Amprenavir

A Look Inside HIV Resistance through Retroviral Protease Interaction Maps

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