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Crystallization and Structural Determination of NRas
Author(s) -
Reid Derion,
Mattos Carla
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.lb202
Subject(s) - neuroblastoma ras viral oncogene homolog , hras , gtpase , allosteric regulation , gtp' , effector , kras , microbiology and biotechnology , guanosine diphosphate , g protein , biology , chemistry , signal transduction , guanosine triphosphate , biochemistry , receptor , mutation , enzyme , gene
NRas is a member of the Ras protein family that is composed of H‐ K‐, and NRas. These small GTPases are involved in signal transduction pathways that control cellular proliferation, apoptosis, motility, survival, and angiogenesis. Each of the Ras superfamily members shares similar structural motifs and function as switches centered on the exchange of guanosine diphosphate (GDP) for guanosine triphosphates (GTP) to activate the switch and on hydrolysis of GTP for deactivation. The overall structure can be divided into a catalytic G domain, which is comprised of the effector lobe (residues 1‐86) and allosteric lobe (residues 87‐166) and the C‐terminal hypervariable region. Once activated, NRas interacts with various effector molecules including Raf, which is essential for cellular growth, and phosphoinositide 3‐kinase (PI3K), which mediates cellular survival. Here, we present the first crystal structure of NRas bound to a non‐hydrolysable GTP analogue, GppNHp, solved to1.67Å resolution from crystal with symmetry of the space group P3 2 21. The structure is compared to NRas‐GDP and to the structures of KRas and HRas bound to GppNHp already in the Protein Data Bank. There are 14 residues that differ in NRas as compared to HRas and KRas. All of these residues are found in the allosteric lobe in areas important for protein‐membrane communication. This research was funded by NSF Grant Number MCB‐1244203.