7NOW
Complex of Nucleoporin-98 and nanobody MS98-27 solved at 1.85A resolution
Summary for 7NOW
| Entry DOI | 10.2210/pdb7now/pdb |
| Descriptor | Anti-Nup98 nanobody MS98-27, Nuclear pore complex protein Nup98, SODIUM ION, ... (5 entities in total) |
| Functional Keywords | nup98, nanobody, complex, nuclear protein |
| Biological source | Vicugna pacos More |
| Total number of polymer chains | 4 |
| Total formula weight | 62837.65 |
| Authors | Sola-Colom, M.,Trakhanov, S.,Goerlich, D. (deposition date: 2021-02-26, release date: 2021-04-07, Last modification date: 2024-10-16) |
| Primary citation | 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 Cited by 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: 38649536DOI: 10.1038/s44318-024-00081-w PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.85 Å) |
Structure validation
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