Knauer D J, Wiley H S, Cunningham D D
J Biol Chem. 1984 May 10;259(9):5623-31.
Because mitogenic stimulation by epidermal growth factor (EGF) requires at least an 8--12-h exposure to commit the majority of the cells to divide, a number of complicating processes must be evaluated in an analysis of the relationship between receptor occupancy and the biological response. During this period, the number of cellular EGF receptors changes and the cells continuously bind, internalize, and degrade the hormone. To deal with these changes, we utilized our previously described steady state model that permits an analysis of EGF interactions with cells under steady state conditions. For these studies we derived and measured a number of steady state rate constants to describe the interaction of 125I-EGF with cultured human and mouse fibroblast-like cells. The validity of these constants was established by demonstrating the close agreement between experimentally determined results and predicted results from computer simulations using these constants. These approaches in conjunction with experiments on the mitogenic stimulation of the cells with EGF led to the conclusion that there is a linear relationship between mitogenic stimulation and EGF receptor occupancy at steady state, with no apparent "spare" EGF receptors.
由于表皮生长因子(EGF)的促有丝分裂刺激需要至少8 - 12小时的暴露才能使大多数细胞进入分裂状态,因此在分析受体占有率与生物学反应之间的关系时,必须评估许多复杂的过程。在此期间,细胞表皮生长因子受体的数量会发生变化,并且细胞会持续结合、内化和降解该激素。为了应对这些变化,我们采用了我们之前描述的稳态模型,该模型允许在稳态条件下分析EGF与细胞的相互作用。对于这些研究,我们推导并测量了许多稳态速率常数,以描述¹²⁵I - EGF与培养的人及小鼠成纤维细胞样细胞的相互作用。通过证明实验确定的结果与使用这些常数进行计算机模拟的预测结果之间的密切一致性,确定了这些常数的有效性。这些方法与用EGF对细胞进行促有丝分裂刺激的实验相结合,得出的结论是,在稳态下,促有丝分裂刺激与EGF受体占有率之间存在线性关系,没有明显的“备用”EGF受体。