Department of Biophysics and Radiation Biology, Semmelweis University, Budapest, Hungary.
MTA-SE Molecular Biophysics Research Group, Hungarian Academy of Sciences, Budapest, Hungary.
Methods Mol Biol. 2020;2112:123-130. doi: 10.1007/978-1-0716-0270-6_9.
Transmembrane proteins include membrane channels, pores, and receptors and, as such, comprise an important part of the proteome, yet our knowledge about them is much less complete than about soluble, globular proteins. An important aspect of transmembrane protein structure is their exact position within the lipid bilayer, a feature hard to investigate experimentally at the atomic level. Here we describe MemBlob, a novel approach utilizing difference electron density maps obtained by cryo-EM studies of transmembrane proteins. The idea behind is that the nonprotein part of such maps carries information on the exact localization of the membrane mimetics used in the experiment and can be used to extract the positional information of the protein within the membrane. MemBlob uses a structural model of the protein and an experimental electron density map to provide an estimation of the surface residues interacting with the membrane.
跨膜蛋白包括膜通道、孔和受体,因此是蛋白质组的重要组成部分,但我们对它们的了解远不如对可溶性球形蛋白的了解完整。跨膜蛋白结构的一个重要方面是它们在脂质双层中的精确位置,这一特性很难在原子水平上通过实验进行研究。在这里,我们描述了 MemBlob,这是一种利用 cryo-EM 研究跨膜蛋白获得的差分电子密度图的新方法。其背后的想法是,此类图谱的非蛋白质部分携带了实验中使用的膜类似物的精确定位信息,可用于提取蛋白质在膜内的位置信息。MemBlob 使用蛋白质的结构模型和实验电子密度图来提供与膜相互作用的表面残基的估计。