High-throughput identification of calcium-regulated proteins across diverse proteomes

作者：Timothy M. Locke, Rose Fields, Hayden Gizinski, George M. Otto, Melissa J. S. MacEwen, Domnița-Valeria Rusnac, Ping He, David M Shechner, Chris D McGann, Matthew D. Berg, Judit Villén, Yasemin Sancak, Devin K. Schweppe · 发表于：Cell Reports · 年份：2024 · DOI：10.1016/j.celrep.2024.114879 · 被引用次数：13 · 研究领域：Advanced Proteomics Techniques and Applications、Ubiquitin and proteasome pathways、Mitochondrial Function and Pathology

Calcium ions play important roles in nearly every biological process, yet whole-proteome analysis of calcium effectors has been hindered by a lack of high-throughput, unbiased, and quantitative methods to identify protein-calcium engagement. To address this, we adapted protein thermostability assays in budding yeast, human cells, and mouse mitochondria. Based on calcium-dependent thermostability, we identified 2,884 putative calcium-regulated proteins across human, mouse, and yeast proteomes. These data revealed calcium engagement of signaling hubs and cellular processes, including metabolic enzymes and the spliceosome. Cross-species comparison of calcium-protein engagement and mutagenesis experiments identified residue-specific cation engagement, even within well-known EF-hand domains. Additionally, we found that the dienoyl-coenzyme A (CoA) reductase DECR1 binds calcium at physiologically relevant concentrations with substrate-specific affinity, suggesting direct calcium regulation of mitochondrial fatty acid oxidation. These discovery-based proteomic analyses of calcium effectors establish a key resource to dissect cation engagement and its mechanistic effects across multiple species and diverse biological processes.