IDENTIFICATION AND QUANTIFICATION OF MATRISOME PROTEINS OF MOUSE KIDNEYS

Abstract

ABSTRACTExtracellular matrix (ECM) is essential for tissue homeostasis. Understanding the matrisome (ECM proteome) composition and mechanisms of ECM control in health and disease is crucial for discovering therapeutic agents and diagnostic tools for inflammatory, fibrotic and cancerous conditions. The challenging obstacle in the ECM analysis is the need to optimise matrisome enrichment methods for different organs, diseases, and species. Currently, there is no optimized protocol nor a publicly available matrisome database for mouse kidneys. This limits the power of murine models in renal diseases and development research. In this study, we comparatively explored the matrisome of healthy C57BL/6 mice using two matrisome extraction methods, including the Millipore Compartment Fractionation (Method-1) and the Sequential Extraction (Method-2) approaches. We examined the efficiency of these methods in matrisome profiling by LC-MS/MS, protein identification and label-free quantification using MaxQuant. As a result of the study, 113 matrisome proteins were identified, including 22 proteins that have not been previously listed in the Matrisome Database (MD). Method-2 allowed identification and quantification of all core and ECM-associated matrisome proteins detected by Method-1 and additionally revealed more core matrisome and ECM-associated proteins. By characterisation of the murine renal matrisome enhanced by our methodological insights, this study provides critically important information for biological and medical kidney research.