Project description:Herein, we explore the druggability of BAG-1S (the smallest isoform) with a specific emphasis on its interaction with C-RAF. We firstly characterize the higher order structure of BAG-1S structure to determine druggable sites by HDX-MS. We then map the BAG-1S:c-Raf interface – identifying a 20 amino acid region likely to interact with c-Raf. Following which, we use mutational analysis of BAG-1S to show specific amino acids that disrupt the BAG-1S:c-Raf interaction, downstream signaling and cell growth. Finally, we develop a BAG-1S interacting peptide (from c-Raf) that disrupts BAG-1 protein:protein interactions (including C/B-RAF, CHIP and HSP70) and modified with a cell-penetrating motif which inhibits the growth of cancer cells with associated signaling inhibition and induction of apoptosis.
Project description:Purpose: The goals of this study are to identify cellular pathways to recover a-syn aggregation-mediated toxicity induced in response to CDC or BAG treatment on PD-iPSC derived mDA neurons. Methods: mRNA profiles of 30-day-old PD-mDA neurons with CDC or BAG treatment for 1 day were generated by deep sequencing, in duplicate, using Illumina HiSeq. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. Conclusions: Our study represents the detailed analysis of mDA neuronal transcriptomes in response to CDC or BAG treatments, with biologic replicates, generated by RNA-seq technology. Our results show that CDC or BAG treatment could rescue the toxcity from a-syn aggregation via purine metabolism / ion transport pathways.