7SCW

KRAS full length wild-type in complex with RGL1 Ras association domain

Summary for 7SCW

• Entry DOI: 10.2210/pdb7scw/pdb
• Descriptor: Isoform 2B of GTPase KRas, Ral guanine nucleotide dissociation stimulator-like 1, MAGNESIUM ION, ... (5 entities in total)
• Functional Keywords: gtpase, ralgef, complex, domain-swap, oncoprotein
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 2
• Total formula weight: 32846.25

Authors

Eves, B.J.,Kuntz, D.A.,Ikura, M.,Marshall, C.B. (deposition date: 2021-09-29, release date: 2022-05-18, Last modification date: 2023-10-25)

Primary citation

Eves, B.J.,Gebregiworgis, T.,Gasmi-Seabrook, G.M.C.,Kuntz, D.A.,Prive, G.G.,Marshall, C.B.,Ikura, M.
Structures of RGL1 RAS-Association Domain in Complex with KRAS and the Oncogenic G12V Mutant.
J.Mol.Biol., 434:167527-167527, 2022

PubMed Abstract: Ral Guanine Nucleotide Dissociation Stimulator Like 1 (RGL1) is a RAS effector protein that activates Ral GTPase by stimulating nucleotide exchange. Most structures of RAS-effector complexes are for the HRAS isoform; relatively few KRAS-effector structures have been solved, even though KRAS mutations are more frequent in human cancers. We determined crystal structures of KRAS/RGL1-RAS-association (RA) domain complexes and characterized the interaction in solution using nuclear magnetic resonance spectroscopy, size-exclusion chromatography combined with multi-angle light scattering and biolayer interferometry. We report structures of wild-type KRAS and the oncogenic G12V mutant in complex with the RA domain of RGL1 at < 2 Å resolution. KRAS/RGL1-RA crystallized as a 1:1 heterodimer, whilst KRAS/RGL1-RA crystallized as a heterotetrameric structure in which RGL1-RA dimerized via domain-swapping the C-terminal beta-strand. Solution data indicated that KRAS and KRAS in complex with RGL1-RA both exist predominantly as 1:1 dimers, while tetramerization occurs through very slow association. Through detailed structural analyses, the distance and angle between RAS α1 helix and RBD/RA α1 helix were found to differ significantly among RAS and RBD/RA complexes. The KRAS/RGL1-RA structures possess some of the largest α1/α1 distances (21.7-22.2 Å), whereas the corresponding distances in previously reported RAS/RAF complexes are significantly shorter (15.2-17.7 Å). Contact map analysis identified unique structural signatures involving contacts between the β1-β2 loop of RA and the α1 helix of RAS, clearly distinguishing the KRAS/RGL1-RA (and other RAS/RA complexes) from RAS/RBD complexes. These results demonstrate that RAS effectors employ an assortment of finely-tuned docking surfaces to achieve optimal interactions with RAS.

PubMed: 35257782
DOI: 10.1016/j.jmb.2022.167527

Experimental method

X-RAY DIFFRACTION (1.98 Å)