5NX0

Structure of Spin-labelled T4 lysozyme mutant L115C-R119C-R1 at room temperature

5NX0 の概要

• エントリーDOI: 10.2210/pdb5nx0/pdb
• 分子名称: Endolysin (2 entities in total)
• 機能のキーワード: nitroxide, spin label, t4 lysozyme, electron paramagnetic resonance, epr, hydrolase
• 由来する生物種: Enterobacteria phage T4
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 18576.35

構造登録者

Gohlke, U.,Loll, B.,Consentius, P.,Mueller, R.,Kaupp, M.,Heinemann, U.,Wahl, M.C.,Risse, T. (登録日: 2017-05-09, 公開日: 2017-07-19, 最終更新日: 2024-01-17)

主引用文献

Consentius, P.,Gohlke, U.,Loll, B.,Alings, C.,Heinemann, U.,Wahl, M.C.,Risse, T.
Combining EPR spectroscopy and X-ray crystallography to elucidate the structure and dynamics of conformationally constrained spin labels in T4 lysozyme single crystals.
Phys Chem Chem Phys, 19:20723-20734, 2017

PubMed Abstract: Electron paramagnetic resonance (EPR) spectroscopy in combination with site-directed spin labeling is used to investigate the structure and dynamics of conformationally constrained spin labels in T4 lysozyme single crystals. Within a single crystal, the oriented ensemble of spin bearing moieties results in a strong angle dependence of the EPR spectra. A quantitative description of the EPR spectra requires the determination of the unit cell orientation with respect to the sample tube and the orientation of the spin bearing moieties within the crystal lattice. Angle dependent EPR spectra were analyzed by line shape simulations using the stochastic Liouville equation approach developed by Freed and co-workers and an effective Hamiltonian approach. The gain in spectral information obtained from the EPR spectra of single crystalline samples taken at different frequencies, namely the X-band and Q-band, allows us to discriminate between motional models describing the spectra of isotropic solutions similarly well. In addition, it is shown that the angle dependent single crystal spectra allow us to identify two spin label rotamers with very similar side chain dynamics. These results demonstrate the utility of single crystal EPR spectroscopy in combination with spectral line shape simulation techniques to extract valuable dynamic information not readily available from the analysis of isotropic systems. In addition, it will be shown that the loss of electron density in high resolution diffraction experiments at room temperature does not allow us to conclude that there is significant structural disorder in the system.

PubMed: 28740983
DOI: 10.1039/c7cp03144k

実験手法

X-RAY DIFFRACTION (1.803 Å)