State Key Laboratory of Proteomics, National Center for Protein Sciences-Beijing, Beijing Proteome Research Center, Beijing Institute of Lifeomics, Beijing 102206, China.
Department of Otolaryngology-Head and Neck Surgery, Shanghai Ninth People's Hospital, Ear Institute, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Translational Medicine on Ear and Nose Diseases, Shanghai 200011, China.
Anal Chem. 2022 May 10;94(18):6809-6818. doi: 10.1021/acs.analchem.2c00581. Epub 2022 Apr 29.
Bioactive peptides play important roles in various biological processes. However, the traditional methods for profiling the peptide-interacting proteins require modifications to the peptide molecules, often leading to false identifications. We found that the interaction between peptide ligands and protein receptors induced significant changes in the abundance of the interacting proteins, which is a signature indicating the interaction and providing complementary information for use in the classical thermal proteome profiling (TPP) technique. Herein, we developed a novel Peptide-ligand-induced Abundance Change of proTeinS (PACTS)-assisted TPP strategy for the identification of peptide-interacting proteins based on the peptide-ligand-induced change in protein abundance. The utility and efficacy of this approach were demonstrated by the identification of the interaction of the protein 3-phosphoinositide-dependent protein kinase 1 (PDPK1) and PDPK1-interacting fragment (PIF) pair and by large-scale profiling of the interacting proteins of PIF. The PACTS-assisted TPP approach was applied to describe the interactome of amyloid beta (Aβ) 1-42 in THP-1 cells and resulted in the identification of 103 interacting proteins. Validation experiments indicated that Aβ1-42 interacted directly with fatty acid synthase and inhibited its enzymatic activity, providing insights into fatty acid metabolic disorders in Alzheimer's disease (AD). Overall, PACTS-assisted TPP is an efficient approach, and the newly identified Aβ-interacting proteins provide rich resources for the research on AD.
生物活性肽在各种生物过程中发挥着重要作用。然而，传统的肽相互作用蛋白分析方法需要对肽分子进行修饰，这往往会导致错误的鉴定。我们发现，肽配体与蛋白受体之间的相互作用会导致相互作用蛋白的丰度发生显著变化，这是相互作用的特征，为经典的热蛋白质组谱分析（TPP）技术提供了补充信息。在此，我们开发了一种新的肽配体诱导蛋白丰度变化分析（PACTS）辅助 TPP 策略，用于基于蛋白丰度变化鉴定肽相互作用蛋白。该方法通过鉴定蛋白 3-磷酸肌醇依赖性蛋白激酶 1（PDPK1）和 PDPK1 相互作用片段（PIF）对以及大规模分析 PIF 的相互作用蛋白，证明了该方法的实用性和功效。PACTS 辅助 TPP 方法被用于描述淀粉样蛋白 β（Aβ）1-42 在 THP-1 细胞中的互作组，并鉴定出 103 个相互作用蛋白。验证实验表明，Aβ1-42 与脂肪酸合酶直接相互作用并抑制其酶活性，为阿尔茨海默病（AD）中脂肪酸代谢紊乱提供了新的见解。总的来说，PACTS 辅助 TPP 是一种有效的方法，新鉴定的 Aβ 相互作用蛋白为 AD 的研究提供了丰富的资源。
