Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, UT, USA.
Calico Life Sciences LLC, South San Francisco, CA, USA.
Science. 2023 Mar 10;379(6636):996-1003. doi: 10.1126/science.abm3452. Epub 2023 Mar 9.
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.
代谢网络相互关联，并影响多种细胞过程。介导这些网络的蛋白质-代谢物相互作用通常亲和力较低，难以系统地发现。我们开发了一种将质谱与平衡透析相结合的方法来系统地发现变构作用（MIDAS），以鉴定这些相互作用。对来自人类碳水化合物代谢的 33 种酶的分析确定了 830 种蛋白质-代谢物相互作用，包括已知的调节剂、底物和产物，以及以前未报道过的相互作用。我们对其中的一部分相互作用进行了功能验证，包括长链酰基辅酶 A 对乳酸脱氢酶同工型的特异性抑制。脂肪酸处理细胞会导致依赖于乳酸脱氢酶同工型表达的丙酮酸-乳酸相互转化的丧失。这些蛋白质-代谢物相互作用可能有助于动态、组织特异性的代谢灵活性，使细胞能够在不断变化的营养环境中生长和存活。
