National Institutes of Health/National Cancer Institute (NIH/NCI)
CA221289
米国
National Institutes of Health/National Heart, Lung, and Blood Institute (NIH/NHLBI)
HL122416
米国
National Institutes of Health/National Heart, Lung, and Blood Institute (NIH/NHLBI)
HL071818
米国
引用
ジャーナル: Sci Adv / 年: 2019 タイトル: Cryo-electron microscopy structure and analysis of the P-Rex1-Gβγ signaling scaffold. 著者: Jennifer N Cash / Sarah Urata / Sheng Li / Sandeep K Ravala / Larisa V Avramova / Michael D Shost / J Silvio Gutkind / John J G Tesmer / Michael A Cianfrocco / 要旨: PIP-dependent Rac exchanger 1 (P-Rex1) is activated downstream of G protein-coupled receptors to promote neutrophil migration and metastasis. The structure of more than half of the enzyme and its ...PIP-dependent Rac exchanger 1 (P-Rex1) is activated downstream of G protein-coupled receptors to promote neutrophil migration and metastasis. The structure of more than half of the enzyme and its regulatory G protein binding site are unknown. Our 3.2 Å cryo-EM structure of the P-Rex1-Gβγ complex reveals that the carboxyl-terminal half of P-Rex1 adopts a complex fold most similar to those of phosphoinositide phosphatases. Although catalytically inert, the domain coalesces with a DEP domain and two PDZ domains to form an extensive docking site for Gβγ. Hydrogen-deuterium exchange mass spectrometry suggests that Gβγ binding induces allosteric changes in P-Rex1, but functional assays indicate that membrane localization is also required for full activation. Thus, a multidomain assembly is key to the regulation of P-Rex1 by Gβγ and the formation of a membrane-localized scaffold optimized for recruitment of other signaling proteins such as PKA and PTEN.
EMPIAR-10285 (タイトル: Cryo-electron microscopy structure of the P-Rex1–G-beta-gamma signaling scaffold Data size: 3.0 TB Data #1: Movie files (.tif) for P-Rex1-Gbg [micrographs - multiframe] Data #2: Micrograph files (.mrc) & CTF log files for P-Rex1-Gbg [micrographs - single frame] Data #3: Extracted particles from Warp for P-Rex1-Gbg [picked particles - single frame - processed])
chain_id: A, residue_range: 622-706, source_name: PDB, initial_model_type: experimental model
精密化
空間: REAL / プロトコル: AB INITIO MODEL / 温度因子: 83
得られたモデル
PDB-6pcv: Single Particle Reconstruction of Phosphatidylinositol (3,4,5) trisphosphate-dependent Rac exchanger 1 bound to G protein beta gamma subunits