Analysis Biomolecular Interactions of NtAGO2 for KRAS by BLI (CAT#: STEM-MB-0195-CJ)

Introduction

Mutations in the RAS gene family account for a large percentage of all known genetic aberrations in cancer. Of the three RAS homologues, KRAS is the most frequently mutated, driving the three most lethal cancer types in the United States: pancreatic ductal adenocarcinoma (PDAC), colorectal cancer, and lung adenocarcinoma. KRAS is a small GTPase that relays mitogenic signals from growth factor receptors at the membrane to the nucleus when bound to GTP. Under physiological conditions, nucleotide cycling on KRAS is accelerated and regulated by guanine exchange factors (GEFs) and GTPase activating proteins (GAPs), which promote GTP loading and hydrolysis, respectively. The KRAS structure is compact, comprised only of a G-domain (residues 1–166) and a hypervariable C-terminal region (residues 167–188) that is important for membrane association.

AGO2 is a large and dynamic protein with four distinct domains connected through linker regions. It performs the final step in miRNA maturation, associates with other proteins to form the RISC complex, and cleaves the miRNA:mRNA duplex. These functions are regulated in a context-specific way through a variety of post-translational modifications. Although AGO2 is independently associated with oncogenesis, it was demonstrated that AGO2 expression was necessary for oncogenic transformation in mutant KRAS-dependent cell lines. The interaction between AGO2 and wild-type KRAS was disrupted following the phosphorylation of AGO2 by the epidermal growth factor receptor (EGFR), allowing KRAS association with activating GEFs such as SOS.