Department of Biochemistry, McGill University, Montréal, Québec, Canada.
Biophys J. 2010 Nov 17;99(10):3327-35. doi: 10.1016/j.bpj.2010.10.001.
Although specific proteins have been identified that regulate the membrane association and facilitate intracellular transport of prenylated Rho- and Rab-family proteins, it is not known whether cellular proteins fulfill similar roles for other prenylated species, such as Ras-family proteins. We used a previously described method to evaluate how several cellular proteins, previously identified as potential binding partners (but not effectors) of K-ras4B, influence the dynamics of K-ras association with the plasma membrane. Overexpression of either PDEδ or PRA1 enhances, whereas knockdown of either protein reduces, the rate of dissociation of K-ras from the plasma membrane. Inhibition of calmodulin likewise reduces the rate of K-ras dissociation from the plasma membrane, in this case in a manner specific for the activated form of K-ras. By contrast, galectin-3 specifically reduces the rate of plasma membrane dissociation of activated K-ras, an effect that is blocked by the K-ras antagonist farnesylthiosalicylic acid (salirasib). Multiple cellular proteins thus control the dynamics of membrane association and intercompartmental movement of K-ras to an important degree even under basal cellular conditions.
虽然已经鉴定出一些特定的蛋白质,它们可以调节被异戊二烯基化的 Rho 和 Rab 家族蛋白的膜结合并促进其细胞内运输,但尚不清楚细胞蛋白是否也可以为其他被异戊二烯基化的物质(如 Ras 家族蛋白)发挥类似的作用。我们使用先前描述的方法来评估几种细胞蛋白(先前被鉴定为 K-ras4B 的潜在结合伴侣(而非效应物))如何影响 K-ras 与质膜结合的动态变化。PDEδ 或 PRA1 的过表达会增强 K-ras 与质膜的解离速率,而这两种蛋白的敲低则会降低 K-ras 与质膜的解离速率。钙调蛋白的抑制同样会降低 K-ras 与质膜的解离速率,在这种情况下,这种作用是针对激活形式的 K-ras 的。相比之下,半乳糖凝集素-3 特异性地降低了激活的 K-ras 从质膜解离的速率,而这种作用被 K-ras 拮抗剂法尼基硫代水杨酸(法尼基硫代水杨酸)阻断。因此,即使在基础细胞条件下,多种细胞蛋白也可以在很大程度上控制 K-ras 与质膜的结合和细胞区间运动的动力学。