Mass spectrometry-based analysis of kindlin phosphorylation, ubiquitination and interactome

Kindlin is an essential activator of integrins and thus indispensable in cell to extracellular matrix (ECM) adhesion. Kindlin abundance and functions are tightly regulated by multiple mechanisms including post-translational modifications (PTMs). Despite the critical roles of kindlin during embryonic development and pathogenesis, the knowledge of kindlin’s PTMs and interactome is far from complete. Here we analyze the phosphorylation, ubiquitination and interacting partners of kindlin in flat, firmly adherent interphase cells and round, weakly attached mitotic cells by immunoprecipitation-based mass spectrometry (IP-MS). The analysis generated potential phosphorylation and ubiquitination sites as well as interactors of kindlin and paved the way to investigate PTMs and binding partners of kindlin.