9OT6

Cryo-EM structure of the PI4KA complex bound to an EFR3 interfering nanobody (F3IN)

9OT6 の概要

• エントリーDOI: 10.2210/pdb9ot6/pdb
• EMDBエントリー: 70826
• 分子名称: Phosphatidylinositol 4-kinase alpha, EFR3 interfering Nanobody (F3IN), Tetratricopeptide repeat protein 7B, ... (4 entities in total)
• 機能のキーワード: pi4ka, ttc7b, fam126a, nanobody, complex, signaling protein
• 由来する生物種: Homo sapiens (human); 詳細
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 768409.29

構造登録者

Shaw, A.L.,Suresh, S.,Yip, C.K.,Burke, J.E. (登録日: 2025-05-26, 公開日: 2025-12-03, 最終更新日: 2025-12-24)

主引用文献

Suresh, S.,Shaw, A.L.,Akintola, D.K.,Lunke, M.,Doerr, S.,Rohilla, P.,Balla, T.,Yip, C.K.,Hansen, S.D.,Cobb, J.A.,Burke, J.E.
Development of an inhibitory TTC7B selective nanobody that blocks EFR3 recruitment of PI4KA.
J.Biol.Chem., 301:110886-110886, 2025

PubMed Abstract: Phosphatidylinositol 4 kinase IIIα (PI4KIIIα/PI4KA) is an essential lipid kinase that plays a critical role in regulating plasma membrane (PM) identity. PI4KA is primarily recruited to the PM through the targeted recruitment by the proteins, EFR3A and EFR3B, which bind to the PI4KA accessory proteins, TTC7 (TTC7A/B) and FAM126 (FAM126A/B). Here, we characterized how both EFR3 isoforms interact with all possible TTC7-FAM126 combinations and developed a nanobody that specifically blocked EFR3-mediated PI4KA recruitment in TTC7B-containing complexes. Most EFR3-TTC7-FAM126 combinations show similar binding affinities, with the exception of EFR3A-TTC7B-FAM126A, which binds with a ∼10-fold higher affinity. Moreover, we showed that EFR3B phosphorylation markedly decreased binding to TTC7-FAM126. Using a yeast display approach, we isolated a TTC7B selective nanobody that blocked EFR3 binding. Cryo-EM and hydrogen deuterium exchange mass spectrometry showed an extended interface with both PI4KA and TTC7B that sterically blocks EFR3 binding. The nanobody caused decreased membrane recruitment both on lipid bilayers and in cells, with decreased PM production of phosphatidylinositol 4-phosphate. Collectively, these findings provide new insights into PI4KA regulation and provide a tool for manipulating PI4KA complexes, which may be valuable for therapeutic targeting.

PubMed: 41197736
DOI: 10.1016/j.jbc.2025.110886

実験手法

ELECTRON MICROSCOPY (3.54 Å)