通过固态NMR光谱学揭示了Natronomonas pharaonis halorhodopsin在其晚期光循环中的结构变化
Xin Zhang1, Hajime Tamaki1, Takashi Kikukawa2
1Institute for Protein Research, Osaka University, Japan.
Biophysical chemistry
|October 6, 2024
概括
研究人员使用固态NMR研究Natronomonas pharaonis halorhodopsin (NpHR),揭示了螺旋D向外运动对于离子吸收至关重要. 这种结构变化对于在这种光遗传工具中创建离子入口路径至关重要.
科学领域:
- 生物物理学的生物物理.
- 结构生物学 结构生物学
- 视觉遗传学 视觉遗传学
背景情况:
- 纳特罗诺莫纳斯法拉尼斯哈洛多普辛 (NpHR) 是一种重要的光驱离子,用于光遗传学.
- 由于晶格分析的挑战,了解NPHR化物吸收机制的结构基础是有限的.
研究的目的:
- 使用固态NMR阐明在化物 (Cl-) 吸收过程中NPHR的结构变化.
- 为了确定关键的残留物和在Cl-运输过程中所涉及的 conformational 动态.
主要方法:
- 固态NMR光谱学用于在近乎生理条件下的Cl-bound和-free状态下分析NPHR.
- 化学转移扰动分析以映射结构变化.
- 在NPHR点突变体上进行光化学分析和DNP增强的固态NMR.
主要成果:
- 耗会引起广泛的结构变化,特别是在螺旋D的细胞外 (EC) 部分.
- 突变分析证实了特定残留物在Cl-吸收过程中的重要性.
- 在Ala165 (A165V) 的突变中断了三元组合和晚期光循环,表明向外螺旋D运动.
结论:
- 在EC部分的螺旋D向外运动对于创建细胞外Cl-入口路径至关重要.
- 这种螺旋运动对于NPHR离子循环和三元体完整性至关重要.
- 这项研究提出了一种NpHR Cl-吸收模型,涉及螺旋体D的形状变化.
相关概念视频
¹H NMR of Conformationally Flexible Molecules: Temporal Resolution
811
At room temperature, the chair conformer of cyclohexane undergoes rapid ring flipping between two equivalent chair conformers at a rate of approximately 105 times per second. These two chair conformers are in equilibrium. The rapid ring flipping results in the interconversion of the axial proton to an equatorial proton and an equatorial to the axial proton. Such interconversions are too rapid and cannot be detected on the NMR timescale. Hence, the NMR spectrometer cannot distinguish between the...
811
¹H NMR of Conformationally Flexible Molecules: Variable-Temperature NMR
1.1K
The axial and equatorial protons in cyclohexane can be distinguished by performing a variable-temperature NMR experiment. In this process, except for one proton, the remaining eleven protons are replaced by deuterium. The deuterium substitution avoids the possible peak splitting caused by the spin-spin coupling between the adjacent protons. The remaining proton flips between the axial and equatorial positions.
1.1K
NMR Spectroscopy Of Amines
8.5K
In proton NMR spectroscopy, primary amines and secondary amines showcase their N–H protons as a broad signal in the chemical shift range between δ 0.5 and 5 ppm. The exact position in this range depends on several factors, including sample concentration, hydrogen bonding, and the type of solvent used. Since amine protons undergo fast proton exchange in solution, the protons are labile and therefore do not participate in any splitting with adjacent protons. Thus, the observed peak is...
8.5K
Channel Rhodopsins
2.5K
Most organisms use photoreceptors to sense and respond to light. Examples of photoreceptors include bacteriorhodopsins and bacteriophytochromes in some bacteria, phytochromes in plants, and rhodopsins in the photoreceptor cells of the vertebral retina. The light-sensitive property of these receptors is because of the bound chromophores, such as bilin in the phytochromes and retinal in the rhodopsins.
Rhodopsins belong to the family of cell surface proteins called G-protein coupled receptors,...
Rhodopsins belong to the family of cell surface proteins called G-protein coupled receptors,...
2.5K
Photoreceptors and Visual Pathways
5.9K
At the molecular level, visual signals trigger transformations in photopigment molecules, resulting in changes in the photoreceptor cell's membrane potential. The photon's energy level is denoted by its wavelength, with each specific wavelength of visible light associated with a distinct color. The spectral range of visible light, classified as electromagnetic radiation, spans from 380 to 720 nm. Electromagnetic radiation wavelengths exceeding 720 nm fall under the infrared category,...
5.9K
¹H NMR: Interpreting Distorted and Overlapping Signals
1.0K
Spin systems where the difference in chemical shifts of the coupled nuclei is greater than ten times J are called first-order spin systems. These nuclei are weakly coupled, and their chemical shifts and coupling constant can generally be estimated from the well-separated signals in the spectrum.
As Δν decreases and the signals move closer, the doublets appear increasingly distorted. The intensities of the inner lines increase at the cost of those of the outer lines as the signals are...
As Δν decreases and the signals move closer, the doublets appear increasingly distorted. The intensities of the inner lines increase at the cost of those of the outer lines as the signals are...
1.0K


