9DJQ

T4 Lysozyme T109H/G113H/D127H/V131H co-crystallized with Cu(II)-NTA

9DJQ の概要

• エントリーDOI: 10.2210/pdb9djq/pdb
• 分子名称: Endolysin, COPPER (II) ION, NITRILOTRIACETIC ACID, ... (9 entities in total)
• 機能のキーワード: hydrolase (o-glycosyl), double histidine mutation, dhis-cu(ii)-nta motif, lysozyme, hydrolase
• 由来する生物種: Tequatrovirus T4
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 19918.36

構造登録者

Chen, E.Y.,Besaw, J.E.,Ernst, O.P. (登録日: 2024-09-06, 公開日: 2026-02-18, 最終更新日: 2026-04-15)

主引用文献

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-, 2026

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: 41794029
DOI: 10.1016/j.str.2026.01.010

実験手法

X-RAY DIFFRACTION (1.28 Å)