Department of Cell and Molecular Biology, Tulane University, New Orleans, LA, USA.
Biomedical Engineering, Tulane University, New Orleans, LA, USA.
Methods Mol Biol. 2022;2418:25-39. doi: 10.1007/978-1-0716-1920-9_2.
Phosphorylation of the serine residues in estrogen receptor (ER) α is important in transcriptional activation. Hence, methods to detect such posttranslational modification events are valuable. We describe, in detail, the analysis of the phosphorylated ERα by electrophoretic separation of proteins and subsequent immunoblotting techniques. In particular, phosphorylation of the ERα is one possible outcome of activation of the putative membrane estrogen receptor (mER), GPR30 or GPER1. Hence, phosphorylation represents a crosstalk event between GPR30 and ERα and may be important in estrogen-regulated physiology.
丝氨酸残基在雌激素受体（ER）α中的磷酸化在转录激活中很重要。因此，检测这种翻译后修饰事件的方法很有价值。我们详细描述了通过蛋白质电泳分离和随后的免疫印迹技术分析磷酸化 ERα 的方法。特别是，ERα 的磷酸化是假定的膜雌激素受体（mER）、GPR30 或 GPER1 激活的一种可能结果。因此，磷酸化代表了 GPR30 和 ERα 之间的串扰事件，并且在雌激素调节的生理中可能很重要。
