c-Cbl-mediated protease-activated receptor 1-induced degradation of Src

• Main Authors: Pei-Yi Lee, 李珮宜
• Other Authors: Hua-Wen Fu
• Format: Others
• Language: en_US
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/47594249891438710417

碩士 === 國立清華大學 === 分子與細胞生物研究所 === 95 === Protease-activated receptor 1 (PAR1) is a G protein-coupled receptor for thrombin. In addition to active Src by G protein, PAR1 recruits Src by ��-arrestin to transduce signals. Src is then sorted to lysosomes with the activated PAR1 for degradation. However, the mechanism by which PAR1-induced degradation of Src and the receptor itself is still unclear. It has been reported that degradation of active Src is mediated by c-Cbl, an ubiquitin E3 ligase. To determine whether c-Cbl mediates the degradation of Src and PAR1 after receptor activation, CHO-K1 cells stably expressing FLAG-tagged PAR1 were transiently transfected with a dominant negative c-Cbl mutant to examine its effect on the degradation of Src and PAR1. I found that stimulation of PAR1 induced degradation of Src. The degradation of Src was blocked by chloroquine, a lysosomal inhibitor. The dominant negative c-Cbl mutant lacking the RING-finger domain (�嵇F c-Cbl) inhibited PAR1-induced degradation of Src and partially inhibited the degradation of PAR1. However, �嵇F c-Cbl did not affect the endocytosis of PAR1. Taken together, these results indicated that c-Cbl mediated lysosomal degradation of Src and PAR1 after the receptor activation.