Investigation of Protein-Protein Interactions by Blue Native-PAGE & Mass Spectrometry

摘要

PPIs are the most fundamental structures that allow to an organelle, cell, tissue, organ or organism to survive, adapt and propagate [1-4]. PPIs are 1) static when proteins form stable protein complexes such as mitochondrial electron transport chain complexes and 2) dynamic or transient, when the interaction between proteins is short-lived and temporary (usually responding to a stimulus and having an end effect). While stable PPIs are easier to investigate, sufficient methods to study transient PPIs still do not exist. One example of transient PPIs is welldemonstrated by signal transduction pathways that are activated by receptor tyrosine kinases (e.g. insulin or Eph receptor pathways) [5,6]. Interaction of receptor tyrosine kinases with their ligands leads to the formation of receptor-ligand complexes and autophosphorylation of the receptor’s kinase domain from the cytoplasmic side, which becomes the docking site for many signaling proteins. In turn, these proteins activate signal transduction pathways, which ultimately trigger a cellular event such as glucose uptake or cytoskeletal remodeling [7-9].