RESUMO
Non-coding RNAs have a role in gene regulation and cellular metabolism control. Metabolism produces metabolites which are small molecules formed during the metabolic process. So far, a direct relationship between metabolites and genes is not fully established; however, pseudogenes and their progenitor genes regulate health and disease states. Other non-coding RNAs also contribute to this regulation at different cellular processes. Accumulation and depletion of metabolites accompany the dynamic equilibrium of health and disease state. In this study, metabolites, their roles in the cell, and the link between metabolites and non-coding RNAs are discussed.
Assuntos
RNA Longo não Codificante , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Regulação da Expressão GênicaRESUMO
BACKGROUND: HSP70 is a survival factor for tumor cells in transformation and in tumor progression as well as in anti-apoptotic response. OBJECTIVE: Several inhibitors targeting HSP70 ATPase function displayed off-target effects, but PES, which targets the HSP70 substrate binding domain, prevents tumor cell survival prominently. However, PES may not bind HSP70 in the absence of nucleotide. This research aimed to design a unique inhibitor molecule that works both in the presence and absence of nucleotides to amplify inhibition. METHODS: A set of chimeric coumarine-pyrazole derivatives were determined by in silico techniques and synthesized to elucidate their inhibitory effects. Cell viability experiments displayed KBR1307 as the most efficient inhibitor. A set of characterization experiments were performed, and the results were compared to that of PES agent. Binding constant, ATP hydrolysis rate, and percent aggregation were determined in the presence and absence of inhibitors. RESULTS: In silico docking experiments showed that only KBR1307 binds the HSP70 substrate binding domain and interacts with cochaperone interface. Binding experiments indicated that KBR1307 binds HSP70 both in the presence and absence of nucleotides, but PES does not. Both inhibitors significantly lower HSP70 ATPase activity and substrate protein disaggregation activity. However, KBR1307 displays a lower IC50 value at the MCF-7 cell line compared to PES. Both inhibitors do not alter HSP70 secondary structure composition and overall stability. CONCLUSION: KBR1307 effectively inhibits HSP70 compared to PES and provides a promising template for novel anticancer drug development.
Assuntos
Antineoplásicos/farmacologia , Cumarínicos/farmacologia , Desenho de Fármacos , Proteínas de Choque Térmico HSP70/antagonistas & inibidores , Pirazóis/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/química , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cumarínicos/química , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Células MCF-7 , Estrutura Molecular , Dobramento de Proteína/efeitos dos fármacos , Pirazóis/química , Relação Estrutura-AtividadeRESUMO
Toxoplasma gondii is ubiquitous obligate intracellular parasite and is one of the most important pathogen for humans and animals. In humans, T. gondii has two life forms: tachyzoites and bradyzoites. Tachyzoites form of T. gondii can cause acute infection, and it is called toxoplasmosis. The development of latent bradyzoites from rapidly growing tachyzoites has been linked to cellular and environmental stresses which are associated with heat shock proteins (Hsps). Hsps play a protective role against stressors. Hsp40 is an important member of Hsp family and T. gondii has 36 predicted Hsp40 family members. Therefore, we studied the cloning and biochemical characterization of the T. gondii RH strain Hsp40 protein-Gok1. Hsp40 prevents protein aggregation and induce refolding. Consequently, Hsp40s may play essential roles in the mechanisms of bradyzoite development and survival in the host organism. Hsp40-Gok1 functional and structural properties may facilitate drug design and protein targeting against toxoplasmosis.