Mapping the dynamic Interactomes of “druggable” membrane proteins: roles in human health and disease (1095.20)

Abstract

Despite extensive research in the past decade, there is a lack of in‐depth understanding of protein networks associated with integral membrane proteins because of their unique biochemical features, enormous complexity and multiplicity. This is a major obstacle to understanding the biology of deregulation of these integral membrane proteins which leads to numerous human diseases, and consequently hinders our development of improved and more targeted therapies to help treat these diseases. To address this challenge, we previously developed an in vivo genetic system, called the Membrane Yeast Two‐Hybrid (MYTH) assay, to identify and characterize protein interactors of all yeast ABC transporters and human receptor tyrosine kinases (RTKs), as well as selected cancer stem cell receptors (CSCRs)5 and G‐protein coupled receptors (GPCRs). However, despite MYTH being a robust technology suitable for mapping the PPIs of a wide‐range of membrane proteins, we have found that a significant percentage of mammalian integral membrane proteins cannot be properly analyzed using this system.To address this, we have recently developed a new variant of MYTH suitable for use in mammalian cells, which we have called the Mammalian Membrane Two‐Hybrid (MaMTH) system. During my talk, I will discuss exciting new findings indicating that MaMTH can detect stimuli (hormone/agonist)‐ and phosphorylation‐dependent PPIs. In addition, I will show that MaMTH allows for monitoring of the phosphorylation states of ErbB‐receptor mutants and drug‐induced activity changes of oncogenic variants of the Epidermal Growth Factor Receptor (EGFR). In conclusion, our study illustrates that MaMTH is a powerful tool for investigating dynamic interactomes of human integral membrane proteins and promises significant contributions to therapeutic research.