Department of Applied Chemistry, Dongduk Women's University, Seoul, Korea.
J Cell Biochem. 2011 Feb;112(2):633-42. doi: 10.1002/jcb.22960.
Temperature sensation initiates from the activation of cellular receptors when the cell is exposed to a decrease in temperature. Here, we applied a phosphoproteome profiling approach to the human lung epithelial cell line BEAS-2B to elucidate cellular cold-responsive processes. The primary aim of this study was to determine which intracellular changes of phosphorylation are accompanied by cold sensation. Eighteen protein spots that exhibited differentially phosphorylated changes in cells were identified. Most of the proteins that were phosphorylated after 5 or 10 min were returned to control levels after 30 or 60 min. Identified proteins were mainly RNA-related (i.e., they were involved in RNA binding and splicing). Temperature (18 and 10°C) stimuli showed homologies that were detected for time course changes in phosphoproteome. The data indicated a time-shift between two temperatures. The phosphorylation of putative cold responsive markers, such as ribosomal protein large P0 and heterochromatin-associated proteins 1, were verified by Western blotting in cells transfected with TRPM8 or TRPA1.
温度感觉始于细胞暴露于温度下降时细胞受体的激活。在这里，我们应用磷酸化蛋白质组学方法研究人肺上皮细胞系 BEAS-2B，以阐明细胞对冷的反应过程。本研究的主要目的是确定哪些细胞内磷酸化变化伴随着冷感觉。鉴定出 18 个蛋白点，这些蛋白点在细胞中表现出不同的磷酸化变化。在 5 或 10 分钟后磷酸化的大多数蛋白质在 30 或 60 分钟后恢复到对照水平。鉴定出的蛋白质主要与 RNA 相关（即，它们参与 RNA 结合和剪接）。温度（18 和 10°C）刺激显示出时间过程中磷酸化蛋白质组变化的同源性。数据表明两个温度之间存在时间差。用转染 TRPM8 或 TRPA1 的细胞通过 Western blot 验证了假定的冷反应标记物（如核糖体蛋白大 P0 和异染色质相关蛋白 1）的磷酸化。
