A comprehensive analysis of screening assays for identifying pyruvate kinase M2 modulators

Moumita Ghosh Chowdhury, Venkatesh Muthukumar, Rudradip Das, Amit Shard
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Abstract

Pyruvate kinase M2 (PKM2) is an essential enzyme in cellular metabolism, playing a key role in regulating glycolysis. It has garnered significant attention in cancer research and is also implicated in the development of various chronic inflammatory conditions. As a result, PKM2 has become a target for drug discovery, with efforts focused on developing inhibitors and activators to modulate its activity. This review discusses various assays used to identify and validate these compounds. Enzyme-based assays help to evaluate PKM2 activity by monitoring substrate conversion. Techniques like dynamic laser light scattering spectrometry and surface plasmon resonance assess the interactions and stability of PKM2 with potential drugs. Cell-based assays measure PKM2 expression levels across different cellular contexts. Additional, methods such as fluorescence titration and mass spectrometry (MS/MS) analyze binding affinity and structural changes of PKM2 upon drug interaction. Metabolic assays, including oxygen consumption rate (OCR) and extracellular acidification rate (ECAR), are employed to evaluate the functional effects of PKM2 modulation on cellular metabolism. This review aims to provide a comprehensive overview of these methodologies to enhance the understanding of PKM2's role and facilitate the discovery of therapeutic agents targeting this important enzyme.
丙酮酸激酶M2调节剂筛选方法的综合分析
丙酮酸激酶M2 (Pyruvate kinase M2, PKM2)是细胞代谢中必需的酶,在糖酵解过程中起关键作用。它在癌症研究中引起了极大的关注,也与各种慢性炎症的发展有关。因此,PKM2已成为药物发现的靶点,人们致力于开发抑制剂和激活剂来调节其活性。本文讨论了用于鉴定和验证这些化合物的各种测定方法。基于酶的检测有助于通过监测底物转化来评估PKM2的活性。动态激光散射光谱和表面等离子体共振等技术评估PKM2与潜在药物的相互作用和稳定性。基于细胞的检测测量PKM2在不同细胞环境中的表达水平。此外,荧光滴定和质谱(MS/MS)等方法分析了药物相互作用时PKM2的结合亲和力和结构变化。代谢测定,包括耗氧量(OCR)和细胞外酸化率(ECAR),被用来评估PKM2调节对细胞代谢的功能影响。本文旨在全面概述这些方法,以增强对PKM2作用的理解,并促进针对这一重要酶的治疗剂的发现。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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