Pavel Mahmud Arif, Lam Clarissa, Kashyap Parul, Salehi-Najafabadi Zahra, Singh Gurpreet, Yu Yong
Department of Biological Sciences, St. John's University, 8000 Utopia Parkway, Queens, NY, 11439, USA.
Methods Mol Biol. 2014;1172:185-92. doi: 10.1007/978-1-4939-0928-5_16.
Cytokines are pleiotropic, low-molecular-weight proteins that regulate the immune responses to infection and inflammation. They stimulate the immune responses by binding to cytokine receptors on the cell plasma membrane. Thus, knowledge of the expression level of particular cytokine receptors on cell surface is crucial for understanding the cytokine function and regulation. One of the techniques to explore the membrane embedded cytokine receptors is cell surface biotinylation. Biotinylated surface proteins can be rapidly purified through the strong interaction between biotin and streptavidin. Here, we describe the procedure for surface biotinylation and purification of biotinylated cytokine receptors for further downstream analysis.
细胞因子是多效性的低分子量蛋白质，可调节对感染和炎症的免疫反应。它们通过与细胞质膜上的细胞因子受体结合来刺激免疫反应。因此，了解特定细胞因子受体在细胞表面的表达水平对于理解细胞因子的功能和调节至关重要。探索膜嵌入细胞因子受体的技术之一是细胞表面生物素化。生物素化的表面蛋白可以通过生物素与链霉亲和素之间的强相互作用快速纯化。在这里，我们描述了细胞表面生物素化和生物素化细胞因子受体纯化的程序，用于进一步的下游分析。
