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TitleCryo-EM Structures of CRAF/14-3-3 and CRAF/14-3-3/MEK1 Complexes.
Journal, issue, pagesJ Mol Biol, Vol. 436, Issue 6, Page 168483, Year 2024
Publish dateFeb 7, 2024
AuthorsDirk Dedden / Julius Nitsche / Elisabeth V Schneider / Maren Thomsen / Daniel Schwarz / Birgitta Leuthner / Ulrich Grädler /
PubMed AbstractRAF protein kinases are essential effectors in the MAPK pathway and are important cancer drug targets. Structural understanding of RAF activation is so far based on cryo-electron microscopy (cryo-EM) ...RAF protein kinases are essential effectors in the MAPK pathway and are important cancer drug targets. Structural understanding of RAF activation is so far based on cryo-electron microscopy (cryo-EM) and X-ray structures of BRAF in different conformational states as inactive or active complexes with KRAS, 14-3-3 and MEK1. In this study, we have solved the first cryo-EM structures of CRAF/14-3-3 at 3.4 Å resolution and CRAF/14-3-3/MEK1 at 4.2 Å resolution using CRAF kinase domain expressed as constitutively active Y340D/Y341D mutant in insect cells. The overall architecture of our CRAF/14-3-3 and CRAF/14-3-3/MEK1 cryo-EM structures is highly similar to corresponding BRAF structures in complex with 14-3-3 or 14-3-3/MEK1 and represent the activated dimeric RAF conformation. Our CRAF cryo-EM structures provide additional insights into structural understanding of the activated CRAF/14-3-3/MEK1 complex.
External linksJ Mol Biol / PubMed:38331211
MethodsEM (single particle)
Resolution3.46 - 4.25 Å
Structure data

16660

8chf

EMDB-16660, PDB-8chf:
cryo-EM Structure of Craf:14-3-3:Mek1
Method: EM (single particle) / Resolution: 4.25 Å

16779

8cpd

EMDB-16779, PDB-8cpd:
Cryo-EM structure of CRaf dimer with 14:3:3
Method: EM (single particle) / Resolution: 3.46 Å

Chemicals

ChemComp-29L:
2-{4-[(1E)-1-(hydroxyimino)-2,3-dihydro-1H-inden-5-yl]-3-(pyridin-4-yl)-1H-pyrazol-1-yl}ethanol

Source
  • homo sapiens (human)
  • spodoptera frugiperda (fall armyworm)
KeywordsSTRUCTURAL PROTEIN / Kinase / CYTOKINE / Complex

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