7NOW

Complex of Nucleoporin-98 and nanobody MS98-27 solved at 1.85A resolution

7NOW の概要

• エントリーDOI: 10.2210/pdb7now/pdb
• 分子名称: Anti-Nup98 nanobody MS98-27, Nuclear pore complex protein Nup98, SODIUM ION, ... (5 entities in total)
• 機能のキーワード: nup98, nanobody, complex, nuclear protein
• 由来する生物種: Vicugna pacos; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 62837.65

構造登録者

Sola-Colom, M.,Trakhanov, S.,Goerlich, D. (登録日: 2021-02-26, 公開日: 2021-04-07, 最終更新日: 2024-10-16)

主引用文献

Sola Colom, M.,Fu, Z.,Gunkel, P.,Guttler, T.,Trakhanov, S.,Srinivasan, V.,Gregor, K.,Pleiner, T.,Gorlich, D.
A checkpoint function for Nup98 in nuclear pore formation suggested by novel inhibitory nanobodies.
Embo J., 2024

PubMed Abstract: Nuclear pore complex (NPC) biogenesis is a still enigmatic example of protein self-assembly. We now introduce several cross-reacting anti-Nup nanobodies for imaging intact nuclear pore complexes from frog to human. We also report a simplified assay that directly tracks postmitotic NPC assembly with added fluorophore-labeled anti-Nup nanobodies. During interphase, NPCs are inserted into a pre-existing nuclear envelope. Monitoring this process is challenging because newly assembled NPCs are indistinguishable from pre-existing ones. We overcame this problem by inserting Xenopus-derived NPCs into human nuclear envelopes and using frog-specific anti-Nup nanobodies for detection. We further asked whether anti-Nup nanobodies could serve as NPC assembly inhibitors. Using a selection strategy against conserved epitopes, we obtained anti-Nup93, Nup98, and Nup155 nanobodies that block Nup-Nup interfaces and arrest NPC assembly. We solved structures of nanobody-target complexes and identified roles for the Nup93 α-solenoid domain in recruiting Nup358 and the Nup214·88·62 complex, as well as for Nup155 and the Nup98 autoproteolytic domain in NPC scaffold assembly. The latter suggests a checkpoint linking pore formation to the assembly of the Nup98-dominated permeability barrier.

PubMed: 38649536
DOI: 10.1038/s44318-024-00081-w

実験手法

X-RAY DIFFRACTION (1.85 Å)