研究人员开发了与平衡透析相结合的质谱技术（MIDAS），用于系统地发现异构体，从而确定这种相互作用。对人类碳水化合物代谢的33种酶的分析确定了830种蛋白质-代谢物的相互作用，包括已知的调节剂、底物和产物以及以前未报告的相互作用。研究人员在功能上验证了一组相互作用，包括长链酰基辅酶A对乳酸脱氢酶的特异性抑制，用脂肪酸处理细胞会导致依赖于乳酸脱氢酶同工酶表达的丙酮酸-乳酸相互转化的损失。这些蛋白质-代谢物的相互作用可能有助于动态的、针对组织的代谢灵活性，使其在不断变化的营养环境中生长和生存。
据了解，代谢网络是相互关联的，影响着不同的细胞过程。介导这些网络的蛋白质-代谢物的相互作用经常是低亲和力的，并且对系统地发现这些作用具有挑战性。
附：英文原文
Title: Protein-metabolite interactomics of carbohydrate metabolism reveal regulation of lactate dehydrogenase
Author: Kevin G. Hicks, Ahmad A. Cluntun, Heidi L. Schubert, Sean R. Hackett, Jordan A. Berg, Paul G. Leonard, Mariana A. Ajalla Aleixo, Youjia Zhou, Alex J. Bott, Sonia R. Salvatore, Fei Chang, Aubrie Blevins, Paige Barta, Samantha Tilley, Aaron Leifer, Andrea Guzman, Ajak Arok, Sarah Fogarty, Jacob M. Winter, Hee-Chul Ahn, Karen N. Allen, Samuel Block, Iara A. Cardoso, Jianping Ding, Ingrid Dreveny, William C. Gasper, Quinn Ho, Atsushi Matsuura, Michael J. Palladino, Sabin Prajapati, Pengkai Sun, Kai Tittmann, Dean R. Tolan, Judith Unterlass, Andrew P. VanDemark, Matthew G. Vander Heiden, Bradley A. Webb, Cai-Hong Yun, Pengkai Zhao, Bei Wang, Francisco J. Schopfer, Christopher P. Hill, Maria Cristina Nonato, Florian L. Muller, James E. Cox, Jared Rutter
Issue&Volume: 2023-03-10
Abstract: Metabolic networks are interconnected and influence diverse cellular processes. The protein-metabolite interactions that mediate these networks are frequently low affinity and challenging to systematically discover. We developed mass spectrometry integrated with equilibrium dialysis for the discovery of allostery systematically (MIDAS) to identify such interactions. Analysis of 33 enzymes from human carbohydrate metabolism identified 830 protein-metabolite interactions, including known regulators, substrates, and products as well as previously unreported interactions. We functionally validated a subset of interactions, including the isoform-specific inhibition of lactate dehydrogenase by long-chain acyl–coenzyme A. Cell treatment with fatty acids caused a loss of pyruvate-lactate interconversion dependent on lactate dehydrogenase isoform expression. These protein-metabolite interactions may contribute to the dynamic, tissue-specific metabolic flexibility that enables growth and survival in an ever-changing nutrient environment.
DOI: abm3452
Source: https://www.science.org/doi/10.1126/science.abm3452