Cell-Derived Vesicles for Single-Molecule Imaging of Membrane Proteins
Cell-Derived Vesicles for Single-Molecule Imaging of Membrane Proteins
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DOI：
10.1002/anie.201408707
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发表时间：
2015-01-07
影响因子：
16.6
通讯作者：
Richards, Christopher I.
中科院分区：
文献类型：
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作者：
Moonschi, Faruk H.;Effinger, Amy K.;Richards, Christopher I.
提出了一种新的方法，通过使用来自细胞表达荧光标记受体的囊泡，应用单分子成像膜受体。在分离囊泡的过程中，受体仍然嵌入在所得囊泡的膜中，从而允许这些囊泡作为用于单分子测量的纳米容器。细胞衍生的囊泡通过将跨膜受体保持在其生理膜中来维持跨膜受体的结构完整性。结果表明，在这些囊泡中分离的受体可以用基于溶液的荧光相关光谱法（FCS）进行研究，并且可以在固体基质上分离用于单分子研究。该技术被应用于确定34烟碱受体的化学计量。该方法提供了将单分子研究扩展到以前无法访问的受体类别的能力。
A new approach is presented for the application of single-molecule imaging to membrane receptors through the use of vesicles derived from cells expressing fluorescently labeled receptors. During the isolation of vesicles, receptors remain embedded in the membrane of the resultant vesicles, thus allowing these vesicles to serve as nanocontainers for single-molecule measurements. Cell-derived vesicles maintain the structural integrity of transmembrane receptors by keeping them in their physiological membrane. It was demonstrated that receptors isolated in these vesicles can be studied with solution-based fluorescence correlation spectroscopy (FCS) and can be isolated on a solid substrate for single-molecule studies. This technique was applied to determine the stoichiometry of 34 nicotinic receptors. The method provides the capability to extend single-molecule studies to previously inaccessible classes of receptors.