Wang X, Xu B, Souza S C, Kopchick J J
Department of Biological Sciences, Ohio University, Athens 45701.
Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1391-5. doi: 10.1073/pnas.91.4.1391.
Porcine and bovine GH receptor (GHR) cDNAs were stably expressed in mouse L cells, which normally do not possess detectable levels of mouse GHR. Expression of the GHR cDNAs resulted in specific binding of 125I-labeled GH by these cell lines. To study GHR-related signaling events in these cells, protein tyrosine phosphorylation was examined. In GH-treated cells, a tyrosine-phosphorylated protein with a molecular mass of approximately 95 kDa (pp95) was increased dramatically (approximately 100-fold) relative to non-GH-treated cells. The amount of pp95 within the cells after GH treatment was positively correlated with the number of GHRs on the cells. Tyrosine phosphorylation of pp95 could not be induced by prolactin, insulin, insulin-like growth factor I, interleukin 2, epidermal growth factor, platelet-derived growth factor, or fibroblast growth factor. Phosphorylation of pp95 was found to be a rapid event that could be observed 60 sec after GH treatment. Also, pp95 appears to exist as a complex of two proteins, i.e., pp95 and pp96. The GH-induced response by these cells may be of use in screening GH analogs for biological activity.
猪和牛的生长激素受体(GHR)cDNA在小鼠L细胞中稳定表达,这些细胞通常不具有可检测水平的小鼠GHR。GHR cDNA的表达导致这些细胞系对125I标记的生长激素具有特异性结合。为了研究这些细胞中与GHR相关的信号转导事件,检测了蛋白质酪氨酸磷酸化。在生长激素处理的细胞中,相对于未用生长激素处理的细胞,一种分子量约为95 kDa的酪氨酸磷酸化蛋白(pp95)显著增加(约100倍)。生长激素处理后细胞内pp95的量与细胞上GHR的数量呈正相关。催乳素、胰岛素、胰岛素样生长因子I、白细胞介素2、表皮生长因子、血小板衍生生长因子或成纤维细胞生长因子均不能诱导pp95的酪氨酸磷酸化。发现pp95的磷酸化是一个快速事件,在生长激素处理后60秒即可观察到。此外,pp95似乎以两种蛋白质的复合物形式存在,即pp95和pp96。这些细胞对生长激素的诱导反应可能有助于筛选具有生物活性的生长激素类似物。