Chirumamilla Chandra Sekhar, Fazil Mobashar Hussain Urf Turabe, Perez-Novo Claudina, Rangarajan Savithri, de Wijn Rik, Ramireddy Padma, Verma Navin Kumar, Vanden Berghe Wim
Laboratory of Protein Chemistry, Proteomics and Epigenetic Signalling (PPES), Department of Biomedical Sciences, University of Antwerp (UA), Antwerpen, Belgium.
Lymphocyte Signalling Research Laboratory, Lee Kong Chain School of Medicine, Nanyang Technological University Singapore, Singapore, Singapore.
Methods Mol Biol. 2019;1930:99-113. doi: 10.1007/978-1-4939-9036-8_13.
T-Lymphocyte kinases are important checkpoints that control T-cell motility by regulating a diverse range of signal transduction pathways. The distinct configuration of kinase events in T-cell could be used to fingerprint the status of T-cells. However, only small fraction human kinases have been characterized so far and little is known about the dynamics of the kinome in motile T-cells. Although several direct and indirect strategies exist to characterize cellular kinase activities, such as RNA interference, antibody arrays, enzyme kinetics, and mass spectrometry, this chapter focuses on an alternative multiplex phosphopeptide array-based methodology, which allows the kinome-wide identification of hyper-activated kinases involved in the regulation of T-cell migration.
T淋巴细胞激酶是重要的检查点，通过调节多种信号转导途径来控制T细胞的运动。T细胞中激酶事件的独特配置可用于描绘T细胞的状态。然而，到目前为止，只有一小部分人类激酶得到了表征，对于运动性T细胞中激酶组的动态变化知之甚少。尽管存在几种直接和间接的策略来表征细胞激酶活性，如RNA干扰、抗体阵列、酶动力学和质谱分析，但本章重点介绍一种基于多重磷酸肽阵列的替代方法，该方法可在全激酶组范围内鉴定参与调节T细胞迁移的过度激活的激酶。
