Zippel R, Morello L, Toschi L, Alberghina L, Comoglio P M, Sturani E
Dipartimento di Fisiologia e Biochimica Generali, Università di Milano, Italy.
Biochim Biophys Acta. 1988 Oct 7;971(3):351-7. doi: 10.1016/0167-4889(88)90151-6.
The conditions affecting the appearance and disappearance of platelet-derived growth factor (PDGF) receptors from the pool of active cell surface-associated receptors were studied. Receptor molecules were revealed in intact Swiss 3T3 fibroblasts by their ability to bind 125I-labeled PDGF and, due to their property to become phosphorylated in tyrosine following ligand binding, by antibodies to phosphotyrosine. PDGF receptor molecules were found to be quite scarce in exponentially growing fibroblasts as compared to quiescent cells. When growing cells were either shifted to a medium containing plasma or received suramin in the culture medium, cell surface-associated PDGF receptors largely increased. This process required about 12 h. Incubation of quiescent cells in serum, but not in plasma, induced a slow decrement of ligand-activatable receptors. In the presence of PDGF the rate of receptor removal from the cell surface was very rapid and was a function of the PDGF concentration. Quiescent cells deprived of cell-surface receptors by incubation with PDGF reexpressed PDGF receptors in about 14 h.
研究了影响血小板衍生生长因子(PDGF)受体在活性细胞表面相关受体库中出现和消失的条件。通过完整的瑞士3T3成纤维细胞结合125I标记的PDGF的能力,以及由于其在配体结合后酪氨酸磷酸化的特性,利用抗磷酸酪氨酸抗体来揭示受体分子。与静止细胞相比,发现指数生长的成纤维细胞中PDGF受体分子相当稀少。当生长的细胞转移到含有血浆的培养基中或在培养基中加入苏拉明时,细胞表面相关的PDGF受体大量增加。这个过程大约需要12小时。在血清中而非血浆中培养静止细胞会导致可被配体激活的受体缓慢减少。在PDGF存在的情况下,受体从细胞表面去除的速度非常快,并且是PDGF浓度的函数。通过与PDGF孵育而缺乏细胞表面受体的静止细胞在大约14小时内重新表达PDGF受体。