Chen Po-Chia, Hub Jochen S
Institute for Microbiology and Genetics, Georg-August-University Göttingen , Justus-von-Liebig weg 11, 37077 Göttingen, Germany.
J Phys Chem Lett. 2015 Dec 17;6(24):5116-21. doi: 10.1021/acs.jpclett.5b02399. Epub 2015 Dec 9.
In experimental studies of solubilized membrane proteins, the detergent corona influences the protein behavior and the resulting measurement. Thus, combinations of experimental techniques with atomistic modeling have been used to resolve corona structural parameters and distributions. Here, we used small-angle X-ray scattering (SAXS) data and molecular dynamics simulations to study a model protein-detergent complex (PDC) consisting of aquaporin-0 and dodecyl-β-maltoside molecules (βDDM). The corona morphology of single snapshots was found to be rough, but it is smooth and compacted in 100-ns-scale ensemble averages. Individual snapshots therefore were unable to accurately represent the ensemble information as captured by experimental SAXS. Mimicking of annular lipids by detergent was also observed. SAXS prediction using different published methods was used to identify optimal βDDM numbers. Explicit-solvent methods predicted best agreement using 290-βDDM PDCs, but implicit-solvent methods gave unclear predictions due to overcompensation by free solvation-layer density parameters. Thus, ensemble-based approaches and physically motivated constraints will help to extract structural information from SAXS data.
在溶解膜蛋白的实验研究中,去污剂冠层会影响蛋白质行为及最终测量结果。因此,已采用实验技术与原子模型相结合的方法来解析冠层结构参数和分布情况。在此,我们利用小角X射线散射(SAXS)数据和分子动力学模拟,研究了一种由水通道蛋白-0和十二烷基-β-麦芽糖苷分子(βDDM)组成的模型蛋白-去污剂复合物(PDC)。发现单个快照的冠层形态粗糙,但在100纳秒尺度的总体平均值中则是平滑且紧密的。因此,单个快照无法准确呈现实验SAXS所捕捉到的总体信息。还观察到去污剂对环状脂质的模拟。使用不同已发表方法进行SAXS预测,以确定最佳βDDM数量。显式溶剂方法预测使用290个βDDM的PDC时吻合度最佳,但隐式溶剂方法由于自由溶剂层密度参数的过度补偿而给出不明确的预测。因此,基于总体的方法和物理驱动的约束将有助于从SAXS数据中提取结构信息。