Shedding Light on the Interaction of Human Anti-Apoptotic Bcl-2 Protein with Ligands through Biophysical and in Silico Studies
Autor: | Ana Rita Otrelo-Cardoso, Alejandro Panjkovich, João Paquete-Ferreira, Teresa Santos-Silva, Jayaraman Muthukumaran, Dmitri I. Svergun, Filipe Freire, Joao Ramos |
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Přispěvatelé: | UCIBIO - Applied Molecular Biosciences Unit, DQ - Departamento de Química |
Jazyk: | angličtina |
Rok vydání: | 2019 |
Předmět: |
0301 basic medicine
BH3 mimetics Protein Conformation Apoptosis Protein-ligand interactions protein-ligand interactions Ligands lcsh:Chemistry Molecular dynamics chemistry.chemical_compound Protein structure lcsh:QH301-705.5 biophysical methods Spectroscopy Sulfonamides Chemistry General Medicine bioinformatics Computer Science Applications Molecular Docking Simulation Proto-Oncogene Proteins c-bcl-2 in silico ddc:540 Protein Binding 030103 biophysics Thermal shift assay Bioinformatics In silico Single-nucleotide polymorphism Molecular Dynamics Simulation Polymorphism Single Nucleotide Biophysical Phenomena Article Catalysis Venetoclax Inorganic Chemistry 03 medical and health sciences SDG 3 - Good Health and Well-being Humans Computer Simulation Bcl-2 Physical and Theoretical Chemistry Molecular Biology Virtual screening Biophysical methods Binding Sites cell apoptosis venetoclax High throughput virtual screening Organic Chemistry Bridged Bicyclo Compounds Heterocyclic 030104 developmental biology lcsh:Biology (General) lcsh:QD1-999 Biophysics Cell apoptosis high throughput virtual screening |
Zdroj: | International journal of molecular sciences 20(4), 860-(2019). doi:10.3390/ijms20040860 Repositório Científico de Acesso Aberto de Portugal Repositório Científico de Acesso Aberto de Portugal (RCAAP) instacron:RCAAP International Journal of Molecular Sciences International Journal of Molecular Sciences, Vol 20, Iss 4, p 860 (2019) Volume 20 Issue 4 |
DOI: | 10.3204/pubdb-2019-03570 |
Popis: | International journal of molecular sciences 20(4), 860 - (2019). doi:10.3390/ijms20040860 Bcl-2 protein is involved in cell apoptosis and is considered an interesting target for anti-cancer therapy. The present study aims to understand the stability and conformational changes of Bcl-2 upon interaction with the inhibitor venetoclax, and to explore other drug-target regions. We combined biophysical and in silico approaches to understand the mechanism of ligand binding to Bcl-2. Thermal shift assay (TSA) and urea electrophoresis showed a significant increase in protein stability upon venetoclax incubation, which is corroborated by molecular docking and molecular dynamics simulations. An 18 °C shift in Bcl-2 melting temperature was observed in the TSA, corresponding to a binding affinity multiple times higher than that of any other reported Bcl-2 inhibitor. This protein-ligand interaction does not implicate alternations in protein conformation, as suggested by SAXS. Additionally, bioinformatics approaches were used to identify deleterious non-synonymous single nucleotide polymorphisms (nsSNPs) of Bcl-2 and their impact on venetoclax binding, suggesting that venetoclax interaction is generally favored against these deleterious nsSNPs. Apart from the BH3 binding groove of Bcl-2, the flexible loop domain (FLD) also plays an important role in regulating the apoptotic process. High-throughput virtual screening (HTVS) identified 5 putative FLD inhibitors from the Zinc database, showing nanomolar affinity toward the FLD of Bcl-2 Published by Molecular Diversity Preservation International, Basel |
Databáze: | OpenAIRE |
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