9DJE
T4 Lysozyme T109H/G113H
Summary for 9DJE
| Entry DOI | 10.2210/pdb9dje/pdb |
| Descriptor | Endolysin, CHLORIDE ION, HEXANE-1,6-DIOL, ... (4 entities in total) |
| Functional Keywords | hydrolase (o-glycosyl), double histidine mutation, dhis motif, lysozyme, hydrolase |
| Biological source | Tequatrovirus T4 |
| Total number of polymer chains | 1 |
| Total formula weight | 19408.28 |
| Authors | Besaw, J.E.,Ernst, O.P. (deposition date: 2024-09-06, release date: 2026-02-18, Last modification date: 2026-04-15) |
| Primary citation | Besaw, J.E.,Reichenwallner, J.,Chen, E.Y.,Hermet-Teesalu, P.,Tregubenko, A.,Kim, K.,Morizumi, T.,Ustav Jr., M.,Ernst, O.P. Structural characterization of the Cu(II)-NTA spin label on alpha-helices by X-ray crystallography and electron paramagnetic resonance. Structure, 34:645-661.e6, 2026 Cited by PubMed Abstract: Site-directed Cu(II)-labelling in pulsed electron paramagnetic resonance (EPR) spectroscopy has demonstrated narrow Cu(II)-Cu(II) distance distributions suitable to resolve subtle protein conformational changes. The high precision derives from a double histidine (dHis) mutation that effectively locks a Cu(II)-nitrilotriacetic acid (Cu(II)-NTA) moiety in place. To date, no structures featuring the dHis-Cu(II)-NTA motif have been resolved. This work presents the atomic-resolution X-ray crystal structures of seven α-helical dHis sites of T4 lysozyme (T4L) in the presence and absence of Cu(II)-NTA. Our research captured the rigid octahedral coordination of the dHis-Cu(II)-NTA complex as well as non-conventional binding modes, which provide valuable insight into dHis site selection. Pulsed EPR experiments on double dHis T4L mutants displayed remarkable agreement to the crystallography-derived distances. This research showcases the rigid configuration of the dHis-Cu(II)-NTA motif, providing geometric constraints that can be leveraged in modeling and molecular dynamics programs to extract protein structural details from EPR experiments. PubMed: 41794029DOI: 10.1016/j.str.2026.01.010 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.62 Å) |
Structure validation
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