VPS35对多巴胺受体D1降解的调控机制研究

Mechanistic study on the degradation of dopamine receptor D1 by VPS35

目的研究囊泡分拣蛋白35(vacuolar protein sorting-35,VPS35)是否影响G蛋白偶联(G proteincoupled)的多巴胺受体D1 (Dopamine receptor D1)的降解过程及其机制。方法依据放线菌酮(cycloheximide,CHX)可以有效抑制生物体内蛋白的合成的原理,我们CHX处理细胞,来研究VPS35对DRD1的降解可能存在的影响。结果过表达VPS35之后,DRD1蛋白的降解速率比对照组加快了。相反的,在下调VPS35水平后,DRD1的降解速率减慢。另外,VPS35能加速DRD1的溶酶体和泛素蛋白酶体降解,但增加溶酶体降解的效果更加显著。结论(1) VPS35能够有效调控DRD1的降解速率;(2) VPS35主要通过溶酶体途径调控DRD1的降解。

Objective To study whether the vacuolar protein sorting-35( VPS35) could effect the process of degradation of G protein coupled receptor dopamine receptor D1( DRD1) and its possible mechanism. Methods Depend on the cycloheximide( CHX) can restrain the synthesis of the proteins of the organism effectively. We handle cells with CHX to study whether VPS35 effects the degradation of DRD1. Results VPS35 accelerates the degradation of DRD1 after over-expressing it in HEK 293 T cells. On the contrary,after knock-down the VPS35 level the rate of degradation of DRD1 was reduced. In addition,the VPS35 could accelerate the degradation of DRD1 through the pathway of lysosome and ubiquitin proteasome when the effect through pathway of lysosome is more significant. Conclusion VPS35 can regulate the degradation of DRD1 effectively mainly through the pathway of lysosome.