Study on the interaction between HIV reverse transcriptase and its non-nucleoside inhibitor nevirapine by capillary electrophoresis

Chen, Wenjing; Li, Wujie; Ling, Xiaomei; Wang, Xiaowei; Liu, Junyi

doi:10.1016/j.jchromb.2010.04.013

Cited by 10 publications

(7 citation statements)

References 22 publications

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“…Several analytical methods have been reported for determination of NEV including high performance liquid chromatography 10, spectrophotometry 11,12, MALDI/ionization time‐of‐flight mass spectrometry 13, micellar electrokinetic chromatography 14, LC‐MS/MS 15, HPTLC 16, capillary electrophoresis 17, and HPLC‐UV 18. Electrochemical methods 19–21 appear as a cost effective, simple and fast alternative to the above, mostly expensive, methods.…”

Section: Introductionmentioning

confidence: 99%

Electrodeposition of Copper Oxide Nanoparticles on Precasted Carbon Nanoparticles Film for Electrochemical Investigation of anti‐HIV Drug Nevirapine

et al. 2015

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This work describes the development of a novel electrochemical sensor based on electrodeposition of copper oxide nanoparticles onto carbon nanoparticle (CNP) film modified electrode for the analysis of the anti‐HIV drug, nevirapine (NEV). The electrochemical experiments were performed using linear sweep and cyclic voltammetry. Atomic force microscopy was applied for surface characterization of the deposited modifier film (CuO‐CNP) on glassy carbon electrode (GCE). No oxidation peak was observed for NEV on the bare GCE, but both CNP‐GCE and CuO‐CNP‐GCE showed a distinctive anodic response towards NEV with considerable enhancement (276‐fold and 350‐fold, respectively) compared to CuO‐GCE. The mechanism of the electrocatalytic process on the modified electrode surface was investigated by cyclic and linear sweep voltammetry at various potential sweep rates and pHs of the buffer solutions. The modified electrode exhibited linear dynamic range in three concentration intervals (0.1–0.8, 1–10 and 10–100 µM) with a detection limit of 66 nM. The stability, reproducibility, and repetitive usability exhibited by the proposed modified electrode are good enough to make it a suitable sensor for the determination of NEV in real samples with complex matrices such as human blood serum.

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Section: Introductionmentioning

confidence: 99%

Electrodeposition of Copper Oxide Nanoparticles on Precasted Carbon Nanoparticles Film for Electrochemical Investigation of anti‐HIV Drug Nevirapine

et al. 2015

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“…A K d value of 0.02 mM has been reported for dissociation of NVP from the ternary RT-P/T-NVP complex (3), whereas a K d value of~30 mM has been reported for dissociation of the RT-NVP binary complex (17), implying a >10 3 -fold, substrate-dependent binding affinity. In sharp contrast to these results, Maga et al (18) reported an NVP-RT dissociation constant of 0.4 mM that is essentially independent of the presence of P/T or P/T-dNTP.…”

Section: Introductionmentioning

confidence: 99%

Protein-Mediated Antagonism between HIV Reverse Transcriptase Ligands Nevirapine and MgATP

Zheng

Mueller

DeRose

et al. 2013

Biophysical Journal

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Nonnucleoside reverse transcriptase inhibitors (NNRTIs) play a central role in the treatment of AIDS, but their mechanisms of action are incompletely understood. The interaction of the NNRTI nevirapine (NVP) with HIV-1 reverse transcriptase (RT) is characterized by a preference for the open conformation of the fingers/thumb subdomains, and a reported variation of three orders of magnitude between the binding affinity of NVP for RT in the presence or absence of primer/template DNA. To investigate the relationship between conformation and ligand binding, we evaluated the use of methionine NMR probes positioned near the tip of the fingers or thumb subdomains. Such probes would be expected to be sensitive to changes in the local environment depending on the fractions of open and closed RT. Comparisons of the NMR spectra of three conservative mutations, I63M, L74M, and L289M, indicated that M63 showed the greatest shift sensitivity to the addition of NVP. The exchange kinetics of the M63 resonance are fast on the chemical shift timescale, but become slow in the presence of NVP due to the slow binding of RT with the inhibitor. The simplest model consistent with this behavior involves a rapid open/closed equilibrium coupled with a slow interaction of the inhibitor with the open conformation. Studies of RT in the presence of both NVP and MgATP indicate a strong negative cooperativity. Binding of MgATP reduces the fraction of RT bound to NVP, as indicated by the intensity of the NVP-perturbed M230 resonance, and enhances the dissociation rate constant of the NVP, resulting in an increase of the open/closed interconversion rate, so that the M63 resonance moves into the fast/intermediate-exchange regime. Protein-mediated interactions appear to explain most of the affinity variation of NVP for RT.

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“…CE detection of microorganisms can be based on the detection of cells [10][11][12][13][14][15][16][17][18], proteins [19][20][21], liposomes [22], or nucleic acids. Among them, nucleic acid detection eliminates many types of false positive and negative results, offering a more precise diagnosis.…”

Section: Diagnosing Infectious Diseases With Cementioning

confidence: 99%

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

Lian

Zhao

2016

Clinical Chemistry and Laboratory Medicine (CCLM)

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Rapid transmission, high morbidity, and mortality are the features of human infectious diseases caused by microorganisms, such as bacteria, fungi, and viruses. These diseases may lead within a short period of time to great personal and property losses, especially in regions where sanitation is poor. Thus, rapid diagnoses are vital for the prevention and therapeutic intervention of human infectious diseases. Several conventional methods are often used to diagnose infectious diseases, e.g. methods based on cultures or morphology, or biochemical tests based on metabonomics. Although traditional methods are considered gold standards and are used most frequently, they are laborious, time consuming, and tedious and cannot meet the demand for rapid diagnoses. Disease diagnosis using capillary electrophoresis methods has the advantages of high efficiency, high throughput, and high speed, and coupled with the different nucleic acid detection strategies overcomes the drawbacks of traditional identification methods, precluding many types of false positive and negative results. Therefore, this review focuses on the application of capillary electrophoresis based on nucleic detection to the diagnosis of human infectious diseases, and offers an introduction to the limitations, advantages, and future developments of this approach.

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Study on the interaction between HIV reverse transcriptase and its non-nucleoside inhibitor nevirapine by capillary electrophoresis

Cited by 10 publications

References 22 publications

Electrodeposition of Copper Oxide Nanoparticles on Precasted Carbon Nanoparticles Film for Electrochemical Investigation of anti‐HIV Drug Nevirapine

Electrodeposition of Copper Oxide Nanoparticles on Precasted Carbon Nanoparticles Film for Electrochemical Investigation of anti‐HIV Drug Nevirapine

Protein-Mediated Antagonism between HIV Reverse Transcriptase Ligands Nevirapine and MgATP

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

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