Multiplex N-terminomics analysis of cardiomyocyte secretome based on MMP-2 and ADAM17 activity using TMT-TAILS quantitative proteomics

Abstract

The role of Matrix Metalloproteinases (MMPs) in the degradation and extracellular matrix remodeling is but one aspect of the role played by this family of proteases that has 27 members. Once activated, the MMPs cleave substrates such as matrix proteins, proinflammatory mediators, and growth factors, which will activate several signaling pathways that depend on MAP kinases, and are important in processes of vascular tonus regulation, cardiac hypertrophy, and response to lesion. It is now known that activation of MMPs by agonists is a fast process that involves PKC, production of oxidants and other MMPs (like membrane-type MMPs), but it is still necessary to know in more detail about the activation/functions of each MMP, specially because of the great therapeutic potential of these metaloenzymes. Furthermore, MMP-7 has been described to increase the concentrations of oxidants, a very important finding, since it changes the paradigm nowadays used to think about the regulation of cardiovascular effects evoked by MMPs and redox processes. In the phase of thematic grant, we propose to perform the subproject 4 to analyze the secretome of cardiomyocytes exposed to ADAM17 or MMP-2 to be able to depict the proteolytic cascades that are specific for each of these two metalloproteinases. The results obtained here will contribute for the advancement of the understanding of the interfaces between redox processes and metalloproteinases, and will also enable the identification of pathways activated specifically by MMP-2 and ADAM17 in the cardiovascular system, with potential implication for the design of future interventions. (AU)