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Structure paper

TitleCryo-EM structure of a RAS/RAF recruitment complex.
Journal, issue, pagesNat Commun, Vol. 14, Issue 1, Page 4580, Year 2023
Publish dateJul 29, 2023
AuthorsEunyoung Park / Shaun Rawson / Anna Schmoker / Byeong-Won Kim / Sehee Oh / Kangkang Song / Hyesung Jeon / Michael J Eck /
PubMed AbstractRAF-family kinases are activated by recruitment to the plasma membrane by GTP-bound RAS, whereupon they initiate signaling through the MAP kinase cascade. Prior structural studies of KRAS with RAF ...RAF-family kinases are activated by recruitment to the plasma membrane by GTP-bound RAS, whereupon they initiate signaling through the MAP kinase cascade. Prior structural studies of KRAS with RAF have focused on the isolated RAS-binding and cysteine-rich domains of RAF (RBD and CRD, respectively), which interact directly with RAS. Here we describe cryo-EM structures of a KRAS bound to intact BRAF in an autoinhibited state with MEK1 and a 14-3-3 dimer. Analysis of this KRAS/BRAF/MEK1/14-3-3 complex reveals KRAS bound to the RAS-binding domain of BRAF, captured in two orientations. Core autoinhibitory interactions in the complex are unperturbed by binding of KRAS and in vitro activation studies confirm that KRAS binding is insufficient to activate BRAF, absent membrane recruitment. These structures illustrate the separability of binding and activation of BRAF by RAS and suggest stabilization of this pre-activation intermediate as an alternative therapeutic strategy to blocking binding of KRAS.
External linksNat Commun / PubMed:37516774 / PubMed Central
MethodsEM (single particle)
Resolution3.9 - 4.3 Å
Structure data

27428

8dgs

EMDB-27428, PDB-8dgs:
Cryo-EM structure of a RAS/RAF complex (state 1)
Method: EM (single particle) / Resolution: 4.3 Å

27429

8dgt

EMDB-27429, PDB-8dgt:
Cryo-EM structure of a RAS/RAF complex (state 2)
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-40253: Truncated Braf/Mek/14-3-3 complex
Method: EM (single particle) / Resolution: 4.1 Å

Chemicals

ChemComp-AGS:
PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER / ATP-gamma-S, energy-carrying molecule analogue*YM

ChemComp-ZN:
Unknown entry

ChemComp-MG:
Unknown entry

ChemComp-LCJ:
5-[(2-fluoro-4-iodophenyl)amino]-N-(2-hydroxyethoxy)imidazo[1,5-a]pyridine-6-carboxamide

ChemComp-GNP:
PHOSPHOAMINOPHOSPHONIC ACID-GUANYLATE ESTER / GppNHp, GMPPNP, energy-carrying molecule analogue*YM / 5'-Guanylyl imidodiphosphate

Source
  • homo sapiens (human)
  • spodoptera exigua (beet armyworm)
KeywordsTRANSFERASE / kinase complex

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