磷酸化
磷
激酶
苏氨酸
信号转导
丝氨酸
细胞生物学
化学
配体(生物化学)
磷酸化级联
受体酪氨酸激酶
生物化学
受体
蛋白质磷酸化
生物
蛋白激酶A
作者
Zunyong Liu,Sen Hou,Ping He
出处
期刊:Methods in molecular biology
日期:2023-11-30
卷期号:: 205-214
标识
DOI:10.1007/978-1-0716-3511-7_15
摘要
Plasma membrane-resident receptor kinases (RKs) are crucial for plants to sense endogenous and exogenous signals in regulating growth, development, and stress response. Upon perception of ligands by the extracellular domain, RKs are usually activated by auto- and/or trans-phosphorylation of the cytoplasmic kinase domain, which in turn phosphorylates downstream substrates to relay the signaling. Therefore, monitoring ligand-induced in vivo phosphorylation dynamics of RKs and their associated proteins provides mechanistic insight into RK activation and downstream signal transduction. Phos-tag specifically binds phosphomonoester dianions of phosphorylated serine, threonine, and tyrosine residues, which enables Phos-tag-containing SDS-PAGE gels to separate phosphorylated proteins from non-phosphorylated form. Here, we describe a detailed method of Mn2+-Phos-tag SDS-PAGE analysis to detect the ligand-induced in vivo phosphorylation of RKs and associated proteins.
科研通智能强力驱动
Strongly Powered by AbleSci AI