Application of ITC-Based Characterization of Thermodynamic and Kinetic Association of Ligands With Proteins in Drug Design

Xu, Yechun

doi:10.3389/fphar.2018.01133

Cited by 45 publications

(56 citation statements)

References 39 publications

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“…Isothermal titration calorimetry studies 54,55 . Liposomes for isothermal titration calorimetry (ITC) studies were prepared by the thin-film method: the desired volume of DOPC (25 mg/ml, CHCl 3 , Avanti Polar Lipids) and CL (5 mg/ml, EtOH, Avanti Polar Lipids) stock solutions were concentrated to dryness under reduced pressure, and the lipid films were resuspended in HEPES buffer (20 mM, pH 7.4) to prepare 50:150 µM CL/DOPC or 200 µM DOPC liposome dispersions.…”

Section: Methodsmentioning

confidence: 99%

Fused isoselenazolium salts suppress breast cancer cell growth by dramatic increase in pyruvate-dependent mitochondrial ROS production

Makrecka-Kūka

Dimitrijevs

Домрачева

et al. 2020

Sci Rep

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The development of targeted drugs for the treatment of cancer remains an unmet medical need. This study was designed to investigate the mechanism underlying breast cancer cell growth suppression caused by fused isoselenazolium salts. The ability to suppress the proliferation of malignant and normal cells in vitro as well as the effect on NAD homeostasis (NAD+, NADH, and NMN levels), NAMPT inhibition and mitochondrial functionality were studied. The interactions of positively charged isoselenazolium salts with the negatively charged mitochondrial membrane model were assessed. Depending on the molecular structure, fused isoselenazolium salts display nanomolar to high micromolar cytotoxicities against MCF-7 and 4T1 breast tumor cell lines. The studied compounds altered NMN, NAD+, and NADH levels and the NAD+/NADH ratio. Mitochondrial functionality experiments showed that fused isoselenazolium salts inhibit pyruvate-dependent respiration but do not directly affect complex I of the electron transfer system. Moreover, the tested compounds induce an immediate dramatic increase in the production of reactive oxygen species. In addition, the isoselenazolothiazolium derivative selectively binds to cardiolipin in a liposomal model. Isoselenazolium salts may be a promising platform for the development of potent drug candidates for anticancer therapy that impact mitochondrial pyruvate-dependent metabolism in breast cancer cells.

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Section: Methodsmentioning

confidence: 99%

Fused isoselenazolium salts suppress breast cancer cell growth by dramatic increase in pyruvate-dependent mitochondrial ROS production

Makrecka-Kūka

Dimitrijevs

Домрачева

et al. 2020

Sci Rep

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“…Quantitative information on inhibitor binding is critical for developing drugs (Su and Xu, 2018) and understanding how enzymes function in living systems (Hulme and Trevethick, 2010). In fact ITC is primarily used to measure these types of host-guest interactions and a quick literature search for "ITC and inhibitor" yields more than 1,300 articles (using Clarivate Analytics Web of Science).…”

Section: Enzyme Inhibitorsmentioning

confidence: 99%

“…In fact ITC is primarily used to measure these types of host-guest interactions and a quick literature search for "ITC and inhibitor" yields more than 1,300 articles (using Clarivate Analytics Web of Science). In a traditional ITC experiment, the enzyme and inhibitor are placed in the sample cell and injection syringe, respectively, and a series of injections are made, while the instrument records the heat released or absorbed by the binding process itself (Su and Xu, 2018). This gives a wealth of thermodynamic information on the interaction, since the Gibbs energy and enthalpy of binding are detected separately, as the shape and overall magnitude of the saturation isotherm, respectively (Sigurskjold, 2000;Tellinghuisen, 2008;Keller et al, 2012;Su and Xu, 2018).…”

Section: Enzyme Inhibitorsmentioning

confidence: 99%

“…In a traditional ITC experiment, the enzyme and inhibitor are placed in the sample cell and injection syringe, respectively, and a series of injections are made, while the instrument records the heat released or absorbed by the binding process itself (Su and Xu, 2018). This gives a wealth of thermodynamic information on the interaction, since the Gibbs energy and enthalpy of binding are detected separately, as the shape and overall magnitude of the saturation isotherm, respectively (Sigurskjold, 2000;Tellinghuisen, 2008;Keller et al, 2012;Su and Xu, 2018). Under favorable circumstances, the kinetics of binding can be measured as well (Burnouf et al, 2012;Piñeiro et al, 2019).…”

Section: Enzyme Inhibitorsmentioning

confidence: 99%

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Enzyme Kinetics by Isothermal Titration Calorimetry: Allostery, Inhibition, and Dynamics

Wang

Moitessier

et al. 2020

Front. Mol. Biosci.

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Isothermal titration calorimetry (ITC) involves accurately measuring the heat that is released or absorbed in real time when one solution is titrated into another. This technique is usually used to measure the thermodynamics of binding reactions. However, there is mounting interest in using it to measure reaction kinetics, particularly enzymatic catalysis. This application of ITC has been steadily growing for the past two decades, and the method is proving to be sensitive, generally applicable, and capable of providing information on enzyme activity that is difficult to obtain using traditional biochemical assays. This review aims to give a broad overview of the use of ITC to measure enzyme kinetics. It describes several different classes of ITC experiment, their strengths and weaknesses, and recent methodological advancements. A summary of applications in the literature is given and several examples where ITC has been used to investigate challenging aspects of enzyme behavior are presented in more detail. These include examples of allostery, where small-molecule binding outside the active site modulates activity. We describe the use of ITC to measure the strength, mode (i.e., competitive, uncompetitive, or mixed), and association and dissociation kinetics of enzyme inhibitors. Further, we provide examples of ITC applied to complex, heterogeneous mixtures, such as insoluble substrates and live cells. These studies exemplify the wide range of problems where ITC can provide answers, and illustrate the versatility of the technique and potential for future development and applications.

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“…Besides thermodynamic information on binding constants, e.g., enzymes, of substrate reactions and inhibitory constants, kinetic data is also of interest. The possibilities are introduced in [71]. Modern ITC instrumentation allows measurement of very small heat powers and provides a tool for biology to study association processes involving liquid membrane proteins, nucleic acids, macromolecular assemblies, and a great variety of ligands.…”

Section: Biomolecular Interaction Analysismentioning

confidence: 99%

Critical assessment of relevant methods in the field of biosensors with direct optical detection based on fibers and waveguides using plasmonic, resonance, and interference effects

Gauglitz

2020

Anal Bioanal Chem

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Direct optical detection has proven to be a highly interesting tool in biomolecular interaction analysis to be used in drug discovery, ligand/receptor interactions, environmental analysis, clinical diagnostics, screening of large data volumes in immunology, cancer therapy, or personalized medicine. In this review, the fundamental optical principles and applications are reviewed. Devices are based on concepts such as refractometry, evanescent field, waveguides modes, reflectometry, resonance and/or interference. They are realized in ring resonators; prism couplers; surface plasmon resonance; resonant mirror; Bragg grating; grating couplers; photonic crystals, Mach-Zehnder, Young, Hartman interferometers; backscattering; ellipsometry; or reflectance interferometry. The physical theories of various optical principles have already been reviewed in detail elsewhere and are therefore only cited. This review provides an overall survey on the application of these methods in direct optical biosensing. The "historical" development of the main principles is given to understand the various, and sometimes only slightly modified variations published as "new" methods or the use of a new acronym and commercialization by different companies. Improvement of optics is only one way to increase the quality of biosensors. Additional essential aspects are the surface modification of transducers, immobilization strategies, selection of recognition elements, the influence of non-specific interaction, selectivity, and sensitivity. Furthermore, papers use for reporting minimal amounts of detectable analyte terms such as value of mass, moles, grams, or mol/L which are difficult to compare. Both these essential aspects (i.e., biochemistry and the presentation of LOD values) can be discussed only in brief (but references are provided) in order to prevent the paper from becoming too long. The review will concentrate on a comparison of the optical methods, their application, and the resulting bioanalytical quality.

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Application of ITC-Based Characterization of Thermodynamic and Kinetic Association of Ligands With Proteins in Drug Design

Cited by 45 publications

References 39 publications

Fused isoselenazolium salts suppress breast cancer cell growth by dramatic increase in pyruvate-dependent mitochondrial ROS production

Fused isoselenazolium salts suppress breast cancer cell growth by dramatic increase in pyruvate-dependent mitochondrial ROS production

Enzyme Kinetics by Isothermal Titration Calorimetry: Allostery, Inhibition, and Dynamics

Critical assessment of relevant methods in the field of biosensors with direct optical detection based on fibers and waveguides using plasmonic, resonance, and interference effects

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