OBJECTIVE: To explore the interaction between BCL2-associated athanogene 5 (BAG5) and Parkin protein,and the regulatory mechanism of BAG5 protein on the level of Parkin protein. METHODS: We performed GST pull-down assay to identify which domain of PINK1 interacted with Parkin, and generated different deletions of BAG5 to identify the domains. Chase time experiment was done to determine the effect of co-regulation of BAG5 protein on the ubiquitination. We further examined the possible interaction between Parkin and PINK1 in 293A cells by co-immunoprecipitation method. RESULTS: BAG5 directly interacted with the Parkin, and all the 4 BAG domains interacted with the Parkin. BAG5 stabilized the Parkin by interfering its degradation via the ubiquitin-mediated proteasomal pathway. CONCLUSION: BAG5 directly interacts with the Parkin, and BAG5 stabilizes the Parkin via the ubiquitin-mediated proteasomal pathway.
OBJECTIVE: To explore the interaction between BCL2-associated athanogene 5 (BAG5) and Parkin protein,and the regulatory mechanism of BAG5 protein on the level of Parkin protein. METHODS: We performed GST pull-down assay to identify which domain of PINK1 interacted with Parkin, and generated different deletions of BAG5 to identify the domains. Chase time experiment was done to determine the effect of co-regulation of BAG5 protein on the ubiquitination. We further examined the possible interaction between Parkin and PINK1 in 293A cells by co-immunoprecipitation method. RESULTS:BAG5 directly interacted with the Parkin, and all the 4 BAG domains interacted with the Parkin. BAG5 stabilized the Parkin by interfering its degradation via the ubiquitin-mediated proteasomal pathway. CONCLUSION:BAG5 directly interacts with the Parkin, and BAG5 stabilizes the Parkin via the ubiquitin-mediated proteasomal pathway.
Authors: Anja Bruchmann; Corinna Roller; Tamara Vanessa Walther; Georg Schäfer; Sara Lehmusvaara; Tapio Visakorpi; Helmut Klocker; Andrew C B Cato; Danilo Maddalo Journal: BMC Cancer Date: 2013-03-01 Impact factor: 4.430