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首页> 外文OA文献 >Incorporation of the nicotinic acetylcholine receptor into planar multilamellar films: characterization by fluorescence and Fourier transform infrared difference spectroscopy.
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Incorporation of the nicotinic acetylcholine receptor into planar multilamellar films: characterization by fluorescence and Fourier transform infrared difference spectroscopy.

机译：烟碱乙酰胆碱受体掺入平面多层膜中：通过荧光和傅里叶变换红外差光谱法表征。

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摘要

A method for preparing thin, planar films of nicotinic acetylcholine receptor (nAChR) membranes that retain the ability to undergo the resting to desensitized state transition and that are suitable for spectroscopic studies has been developed. Native, alkaline-extracted nAChR membranes from Torpedo are dried under nitrogen on either a plastic microscope coverslip or a germanium internal reflection element (IRE) and then equilibrated with buffer. The drying procedure has no effect on the functional state of the nAChR as judged by a fluorescence assay using the probe ethidium bromide. The times required for an acetylcholine analogue (carbamylcholine), a local anesthetic (dibucaine), and a fluorescent probe (ethidium bromide) to penetrate films of varying degrees of thickness, interact with the receptor, and then to be washed from the films have been established. Under these conditions, the nAChR films can be repetitively cycled between the resting and desensitized states. Both fluorescence and infrared spectroscopy show that the films adhere strongly to either support even with buffer flowing continuously past the film surface. Fourier transform infrared difference spectra calculated from spectra recorded in the presence and absence of carbamylcholine show small, reproducible bands which reflect changes in nAChR structure upon desensitization.
机译：已经开发出一种制备烟碱样乙酰胆碱受体（nAChR）膜的平面薄膜的方法，该膜保持经历从静止到脱敏状态转变的能力并且适合于光谱研究。将Torpedo的天然碱性提取的nAChR膜在氮气下在塑料显微镜盖玻片或锗内部反射元件（IRE）上干燥，然后用缓冲液平衡。干燥过程对nAChR的功能状态没有影响，通过使用溴化乙锭探针的荧光分析可以判断。乙酰胆碱类似物（氨基甲酰胆碱），局麻药（地布卡因）和荧光探针（溴化乙锭）穿透不同厚度的膜，与受体相互作用然后从膜上洗去所需的时间成立。在这些条件下，nAChR膜可以在静止和脱敏状态之间反复循环。荧光和红外光谱都表明，即使缓冲液不断流过薄膜表面，薄膜也可以牢固地粘附在任一支撑物上。由在存在和不存在氨基甲胆碱的情况下记录的光谱计算出的傅立叶变换红外差光谱显示出小的可再现谱带，反映了脱敏后nAChR结构的变化。

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