enzyme kinetics; inhibitor IC50; thermodynamics; vitality value; HIV Protease Inhibitors; HIV Protease; p16 protease, Human immunodeficiency virus 1; Biocatalysis/drug effects; Drug Resistance, Viral/genetics; HIV Protease/chemistry; HIV Protease/genetics; HIV Protease/metabolism; HIV Protease Inhibitors/chemistry; HIV Protease Inhibitors/pharmacology; HIV-1/drug effects; HIV-1/enzymology; HIV-1/genetics; Kinetics; Models, Molecular; Thermodynamics; Mutation; Biocatalysis; Drug Resistance, Viral; HIV-1; Biochemistry; Molecular Biology; Clinical Biochemistry
Abstract :
[en] The efficacy of HIV-1 protease (PR) inhibition therapies is often compromised by the emergence of mutations in the PR molecule that reduces the binding affinity of inhibitors while maintaining viable catalytic activity and affinity for natural substrates. In the present study, we used a recombinant HIV-1 C-SA PR and a recently reported variant for inhibition (Ki, IC50) and thermodynamic studies against nine clinically used inhibitors. This is the first time that binding free energies for C-SA PR and the mutant are reported. This variant PR harbours a mutation and insertion (I36T↑T) at position 36 of the C-SA HIV-1 PR, and did not show a significant difference in the catalytic effect of the HIV-1 PR. However, the nine clinically approved HIV PR drugs used in this study demonstrated weaker inhibition and lower binding affinities toward the variant when compared to the wild type HIV-1 PR. All the protease inhibitors (PIs), except Amprenavir and Ritonavir exhibited a significant decrease in binding affinity (p<0.0001). Darunavir and Nelfinavir exhibited the weakest binding affinity, 155- and 95-fold decreases respectively, toward the variant. Vitality values for the variant PR, against the seven selected PIs, confirm the impact of the mutation and insertion on the South African HIV-1 subtype C PR. This information has important clinical implications for thousands of patients in Sub-Saharan Africa.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Maseko, Sibusiso Bonginkhost ; Catalysis and Peptide Research Unit, School of Health Sciences, University of KwaZulu-Natal, Durban, South Africa
Padayachee, Eden; Catalysis and Peptide Research Unit, School of Health Sciences, University of KwaZulu-Natal, Durban, South Africa
Govender, Thavendran ; Catalysis and Peptide Research Unit, School of Health Sciences, University of KwaZulu-Natal, Durban, South Africa
Sayed, Yasien; Protein Structure-Function Research Unit, School of Molecular and Cell Biology, University of the Witwatersrand, Johannesburg, South Africa
Kruger, Gert ; Catalysis and Peptide Research Unit, School of Health Sciences, University of KwaZulu-Natal, Durban, South Africa
Maguire, Glenn E M ; Catalysis and Peptide Research Unit, School of Health Sciences, University of KwaZulu-Natal, Durban, South Africa ; School of Chemistry and Physics, University of KwaZulu-Natal, Durban, South Africa
Lin, Johnson ; School of Life Sciences, University of KwaZulu-Natal, Durban, South Africa
Language :
English
Title :
I36T↑T mutation in South African subtype C (C-SA) HIV-1 protease significantly alters protease-drug interactions.
Acknowledgements: We thank the NRF, UKZN, University of the Witwatersrand, Aspen Pharmacare and MRC (SA) for financial support. The protease sequence was supplied by Professor Lynn Morris (HIV Research, National Institute for Communicable Diseases, South Africa).
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