Meyerson Joel R, Rao Prashant, Kumar Janesh, Chittori Sagar, Banerjee Soojay, Pierson Jason, Mayer Mark L, Subramaniam Sriram
Laboratory of Cell Biology, Center for Cancer Research, NCI, NIH, Bethesda, MD 20892.
Laboratory of Cellular and Molecular Neurophysiology, Porter Neuroscience Research Center, NICHD, NIH, Bethesda MD 20892.
Sci Rep. 2014 Nov 18;4:7084. doi: 10.1038/srep07084.
Poor partitioning of macromolecules into the holes of holey carbon support grids frequently limits structural determination by single particle cryo-electron microscopy (cryo-EM). Here, we present a method to deposit, on gold-coated carbon grids, a self-assembled monolayer whose surface properties can be controlled by chemical modification. We demonstrate the utility of this approach to drive partitioning of ionotropic glutamate receptors into the holes, thereby enabling 3D structural analysis using cryo-EM methods.
大分子在多孔碳支持网格的孔中分配不佳，这常常限制了单颗粒冷冻电子显微镜（cryo-EM）的结构测定。在此，我们提出一种方法，可在涂金的碳网格上沉积一种自组装单分子层，其表面性质可通过化学修饰来控制。我们展示了这种方法在驱动离子型谷氨酸受体进入孔中分配方面的效用，从而能够使用冷冻电子显微镜方法进行三维结构分析。
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