4FJ3

14-3-3 isoform zeta in complex with a diphoyphorylated C-RAF peptide

4FJ3 の概要

• エントリーDOI: 10.2210/pdb4fj3/pdb
• 関連するPDBエントリー: 1A4O 3IQJ 3NKX
• 分子名称: 14-3-3 protein zeta/delta, RAF proto-oncogene serine/threonine-protein kinase (3 entities in total)
• 機能のキーワード: 14-3-3 fold, all alpha-helical, adapter protein, protein binding-transferase complex, protein binding/transferase
• 由来する生物種: Homo sapiens (human); 詳細
• 細胞内の位置: Cytoplasm: P63104 P04049
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 57624.64

構造登録者

Ottmann, C.,Molzan, M. (登録日: 2012-06-11, 公開日: 2012-09-05, 最終更新日: 2013-07-10)

主引用文献

Molzan, M.,Ottmann, C.
Synergistic binding of the phosphorylated S233- and S259-binding sites of C-RAF to one 14-3-3zeta dimer
J.Mol.Biol., 423:486-495, 2012

PubMed Abstract: C-RAF kinase is a central component of the Ras-RAF-MEK (mitogen-activated protein kinase/extracellular signal-regulated kinase)-ERK (extracellular signal-regulated kinase) pathway, which has been shown to be activated in 30% of human tumors. 14-3-3 proteins inactivate C-RAF by binding to the two N-terminal phosphorylation-dependent binding sites surrounding S233 and S259. 14-3-3 proteins can bind two target sequences located on one polypeptide chain simultaneously, thereby increasing binding affinity compared to single-site binding and possibly allowing regulated 14-3-3 binding through gatekeeper phosphorylation. To date, it was unclear whether 14-3-3 proteins can bind the two N-terminal phosphorylation-dependent binding sites of C-RAF simultaneously. Fluorescence polarization using phosphorylated peptides demonstrated that S233 is the low-affinity and S259 is the high-affinity binding site, while simultaneous engagement of both sites by 14-3-3ζ enhances affinity compared to single-site binding. Determination of a 1:1 stoichiometry for the di-phosphorylated peptide binding to one 14-3-3ζ dimer with isothermal titration calorimetry was supported by the crystal structure of the 14-3-3ζ/C-RAFpS233,pS259 complex. Cellular localization studies validate the significance of these sites for cytoplasmic retention of C-RAF, suggesting an extended mechanism of RAF regulation by 14-3-3 proteins.

PubMed: 22922483
DOI: 10.1016/j.jmb.2012.08.009

実験手法

X-RAY DIFFRACTION (1.95 Å)