9M8W
NMR structure of ProteinMPNN-desighed ubiquitin variant R4 at pH 3 with 8 M urea
Summary for 9M8W
| Entry DOI | 10.2210/pdb9m8w/pdb |
| Descriptor | ProteinMPNN-designed ubiquitin variant R4 (1 entity in total) |
| Functional Keywords | ubiquitin variant, r4, urea, de novo protein |
| Biological source | Escherichia coli BL21(DE3) |
| Total number of polymer chains | 1 |
| Total formula weight | 9439.81 |
| Authors | Wu, K.-P.,Chen, L.-Y.,Chuang, W.-C. (deposition date: 2025-03-13, release date: 2026-02-11, Last modification date: 2026-03-11) |
| Primary citation | Chen, L.Y.,Lu, W.L.,Pathania, T.,Chu, I.H.,Ho, M.R.,Chuang, W.C.,Lou, Y.C.,Hung, T.I.,Miyanoiri, Y.,Chang, C.A.,Wu, K.P. Mesostructured Water Enhances Stability of ProteinMPNN-Designed Ubiquitin-Fold Proteins. J.Am.Chem.Soc., 148:7363-7377, 2026 Cited by PubMed Abstract: AI-designed protein variants have demonstrated remarkable resistance to heat and chemical stress, yet the molecular mechanisms underlying this stability remain unclear. Here, we present a comprehensive biophysical and nuclear magnetic resonance (NMR) analysis of thermally stable ubiquitin and its ProteinMPNN-designed variants, R4 and R10, together with a second system based on the less stable ISG15 C-terminal domain (ISG15-CTD). Both R4/R10 and ProteinMPNN-designed ISG15-CTD variants (ICVs) exhibit extraordinary thermostability beyond 120 °C, and resist extreme denaturation at pH 3.0 in 8 M urea. NMR relaxation and hydrogen-deuterium exchange, and molecular-dynamics simulations reveal a protective mesostructured hydration shell that strengthens the hydrogen bonding network between protein-bound and bulk water, thereby suppressing unfolding. Sequence and electrostatic analyses indicate that this hydration arises from charge enrichment and clustering on the protein surface. These findings identify mesostructured hydration as a general, sequence-encoded mechanism of ProteinMPNN-driven stability and provide a physical framework for designing highly resilient biomolecules. PubMed: 41643123DOI: 10.1021/jacs.5c19875 PDB entries with the same primary citation |
| Experimental method | SOLUTION NMR |
Structure validation
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