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利用弹性非相干中子散射技术研究在生理温度下光系统 II 膜的动态特性。与放氧复合体失活相关的膜的柔韧性增加。

Dynamic properties of photosystem II membranes at physiological temperatures characterized by elastic incoherent neutron scattering. Increased flexibility associated with the inactivation of the oxygen evolving complex.

机构信息

Institut Laue-Langevin, P.O. Box 156, 38042, Grenoble Cedex 9, France.

出版信息

Photosynth Res. 2012 Mar;111(1-2):113-24. doi: 10.1007/s11120-011-9701-x. Epub 2011 Nov 4.

Abstract

Elastic incoherent neutron scattering (EINS), a non-invasive technique which is capable of measuring the mean square displacement of atoms in the sample, has been widely used in biology for exploring the dynamics of proteins and lipid membranes but studies on photosynthetic systems are scarce. In this study we investigated the dynamic characteristics of Photosystem II (PSII) membrane fragments between 280 and 340 K, i.e., in the physiological temperature range and in the range of thermal denaturation of some of the protein complexes. The mean square displacement values revealed the presence of a hydration-sensitive transition in the sample between 310 and 320 K, suggesting that the oxygen evolving complex (OEC) plays an important role in the transition. Indeed, in samples in which the OEC had been removed by TRIS- or heat-treatments (323 and 333 K) no such transition was found. Further support on the main role of OEC in these reorganizations is provided by data obtained from differential scanning calorimetry experiments, showing marked differences between the untreated and TRIS-treated samples. In contrast, circular dichroism spectra exhibited only minor changes in the excitonic interactions below 323 K, showing that the molecular organization of the pigment-protein complexes remains essentially unaffected. Our data, along with earlier incoherent neutron scattering data on PSII membranes at cryogenic temperatures (Pieper et al., Biochemistry 46:11398-11409, 2007), demonstrate that this technique can be applied to characterize the dynamic features of PSII membranes, and can be used to investigate photosynthetic membranes under physiologically relevant experimental conditions.

摘要

弹性非相干中子散射(EINS)是一种非侵入性技术,能够测量样品中原子的均方位移,已广泛应用于生物学领域,用于研究蛋白质和脂质膜的动力学,但在光合作用系统方面的研究较少。在这项研究中,我们研究了 280 至 340 K 之间的 PSII 膜片段的动态特性,即生理温度范围内和一些蛋白质复合物热变性的范围内。均方位移值表明,在 310 至 320 K 之间的样品中存在一个水敏性转变,表明氧释放复合物(OEC)在该转变中起重要作用。事实上,在经过 TRIS 或热处理(323 和 333 K)去除 OEC 的样品中,没有发现这种转变。差示扫描量热法实验获得的数据进一步支持了 OEC 在这些重排中的主要作用,结果表明未经处理和 TRIS 处理的样品之间存在明显差异。相比之下,圆二色光谱在 323 K 以下的激子相互作用中仅显示出较小的变化,表明色素-蛋白复合物的分子组织基本不受影响。我们的数据以及早期在低温下对 PSII 膜的非相干中子散射数据(Pieper 等人,Biochemistry 46:11398-11409, 2007)表明,该技术可用于表征 PSII 膜的动态特征,并可用于在生理相关的实验条件下研究光合作用膜。

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