Proteomics and crystal structure analysis to elucidate vascular pathological mechanisms

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This project was undertaken to elucidate factors important in vascular disease pathophysiology through proteomic methods. Crystal structure analysis was also used to understand the physiochemical basis of the mechanisms of action. These studies were done with the ultimate objective to make possible the development of new therapies and diagnostic methods.KLF5 is a factor which regulates the cardiovascular remodeling process in response to stress, and is presently considered one of the most important cardiovascular pathological factors. To understand its regulation, interacting proteins were identified by proteomic methods, which included the repressor SET. The functions of SET on KLF5 were analyzed. The crystal structure of SET was also solved which will allow for 'pin-point' drug design.We also showed that KLF5 is acetylated, and for p300 to be the acetylase while SET inhibits this acetylation. Positive and negative regulation of KLF5 by respective p300 and SET factors was coupled to interaction and acetylation. The deacetylase HDAC1 further negatively regulated KLF5, and to physically compete with p300. KLF5 was also shown to possess anti-apoptotic activity, through analysis of which, we showed that KLF5 interacts with a pro-apoptotic fragment of PARP-1, and for this interaction to also be regulated by acetylation of KLF5.Through our studies, we have shown how interaction and modifications functionally affect the actions of KLF5, with a particular focus on epigenetic regulation. This better understanding of molecular actions in vascular pathology will allow for development of new therapies.