Structural charaterization of RNA binding proteins and fragment-based screening by Liquid Chromatography and mass spectrometry

Abstract

With the aim of expanding the druggable proteome and lead to the discovery of new medicines, we work on the characterization of understudied human proteins, which show preliminary connection to disease. Within this aim, research has been expanded on the importance of protein related to RNA biology as antibody and drug targets, including the proteins 'per se' and their upstream modulators as kinases and methyltranferases. Thus, RNA binding proteins as well as kinase proteins constitute important biological targets for the advancement of biopharma. Those proteins can be targeted by small-molecule chemical probes, which are molecules defined as having high affinity and selectivity for a target and which can enter the cell to engage the target in vivo. Probes work as a complement to biological reagents for target validation and can be refined to become new drugs. In addition, biological targets can also lead to immunotherapy. In this project we aim to contribute for the characterization of the RNA binding protein BICC (Bicaudal C Homology 1) and UHMK (U2AF homology motif Serine/Threonine-Protein kinase) as potential drug targets related to cancer and neurological. We will characterize its interactors, target mRNAs and search for new chemical ligands.- understand the RPBs function in relation to the binding of RNA by determining its protein interactome, using immunoprecipitation followed by mass spectrometry (IP-MS). - elucidate which bound mRNAs can clarify a link to disease, using crosslinking immunoprecipitation followed by RNA-seq (CLIP-seq). - understand how the RBPs fold and bind to target mRNAs, by combining synthetic sequences of RNA binding motifs from the CLIP-seq with the recombinant protein, using co-crystallography; - suggest small molecules to be used as chemical starting points for mapping the protein chemical space and for leading to inhibitors, using fragment-based lead discovery and determination by mass spectrometry. (AU)