Interaction between hHBrk1 and WAVE1 protein

AIM: To identify the binding sites between hHBrk1 and WAVE1 protein.METHODS: PCR was applied to clone the WAVE1 gene and its truncated domains from human fetal brain library. The gene fragments were sub-cloned into eukaryotic expression vector. Co-immunoprecipitation assay was used to identify the binding sites between hHBrk1 and WAVE1.RESULTS: The sequences of WAVE1 and the truncated domains were successfully amplified and sub-cloned into pCMV-HA vector. An HA tag was added to the fused protein at N-terminal. The interaction between hHBrk1 and the Scar homology domain(SHD) of WAVE1 was identified by co-immunoprecipitation. We also observed that WAVE1 bound to the heptad repeat(HR) domain of hHBrk1, and the site-directed mutations at HR domain abolished the interaction between hHBrk1 and WAVE1.CONCLUSION: hHBrk1 may play an important role in migration of tumor cells through binding to SHD of WAVE1 protein.