In vitro study of drug-protein interaction using electronic absorption, fluorescence, and circular dichroism spectroscopy

Document Type : Original Articles

Authors

1 Department of Biology, Faculty of Food Industry & Agriculture, Standard Research Institute, Karaj, Iran

2 Department of Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Karaj, Iran

Abstract

In the near future, design of a new generation of drugs targeting proteins will be required. Considering the complex bond between the drug and protein, the structure and stability of the target protein should be considered. So far, a series of in vitro investigations have been conducted with the aim of predicting drug-biological medium interactions. In these studies, use of spectroscopic methods, such as electronic absorption, fluorescence, and circular dichroism spectroscopy, which are briefly discussed in the present study, has been highlighted. The binding affinity of drug(s) to protein(s) and their binding mechanism(s) can be clearly determined by these methods, which reveal reactions in biological systems at low concentrations under physiological conditions. Ultraviolet-visible spectroscopy can be used as an accessible tool to measure slight changes in protein structure. Moreover, fluorescence spectroscopy provides tertiary structural information. On the other hand, circular dichroism spectroscopy in far-ultraviolet regions (180–260 nm) yields suitable information about different secondary structures of proteins. Conformational changes of proteins due to alterations such as physicochemical conditions, in vitro chemical modifications, and drug binding could impact ultraviolet-visible absorption, circular dichroism, and fluorescence spectra. Therefore, the study of changed spectra could reveal the structure-activity relationship of drug compounds and target proteins. In the present study, a short description of the mentioned methods, along with some related equations which are usually used to analyze and discuss the preliminary data, is presented. Overall, the obtained results could facilitate the development of biological and pharmaceutical potentials of drugs in the future.

Keywords

Main Subjects

Article Title [French]

Etude in Vitro de l’interaction médicament-protéine par absorption électronique, fluorescence et dichroïsme circulaire

Abstract [French]

Dans un futur proche, la conception d’une nouvelle génération de médicaments ciblant des protéines sera nécessaire. Etant donnée la complexité des liaisons entre médicaments et protéines, la structure et la stabilité de la protéine cible doivent également être prises en compte. Jusqu’alors, une série d’études in vitro ont été menées dans le but de prédire les interactions potentielles entre médicament et milieu biologique. Dans cette étude, plusieurs méthodes spectroscopiques comme l’absorption électronique, la fluorescence et le dichroïsme circulaire ont été utilisées dans ce sens. Ces méthodes sont capables de révéler, dans des conditions physiologiques et dans de faibles concentrations, l’affinité des liaisons médicaments-protéines survenant dans des systèmes biologiques variés. De plus, la spectroscopie à l’ultraviolet visible représente une technique accessible pour mesurer de légers changements structurels dans les protéines d’intérêts. Des données concernant les structures secondaires et tertiaires des protéines ont été respectivement obtenues par spectroscopie à ultraviolet lointain (180–260 nm) et spectroscopie à fluorescence. Notre étude montre que les changements de conformation des protéines induites par des altérations physicochimiques, des modifications chimiques in vitro ou des liaisons avec des composés médicamenteux peuvent avoir un impact sur les spectres obtenus par absorption à ultraviolet-visible, par dichroïsme circulaire ou par fluorescence. Par conséquent, l’étude des spectres modifiés peut révéler la relation structure-activité entre composés médicamenteux et protéines cibles. Les différentes méthodes de spectrométries utilisées ont été brièvement décrites et les équations habituellement utilisées pour analyser et développer les résultats préliminaires sont présentées. Dans leur globalité, les résultats obtenus peuvent à l’avenir faciliter le développement de produits biologiques et pharmaceutiques.

Keywords [French]

  • Etude in vitro
  • Interaction médicament-protéine
  • Structure des protéines
  • Spectroscopie
  • Absorption électronique
  • Fluorescence
  • Dichroïsme circulaire

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Archives of Razi Institute
Volume 71, Issue 3
September and October 2016
Pages 183-194

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