Hess G P, Aoshima H, Cash D J, Lenchitz B
Proc Natl Acad Sci U S A. 1981 Mar;78(3):1361-5. doi: 10.1073/pnas.78.3.1361.
The specific reaction rate (J) of the acetylcholine receptor-controlled ion translocation has been determined. In eel Ringer's solution (pH 7.0) at 1 degrees C, J = 3 X 10(7) M-1 sec-1. J is an intrinsic constant that is characteristic of the receptor and independent of other properties of a receptor-containing cell that also determine the rates of ion translocation. Membrane vesicles (prepared from the electric organ of Electrophorus electricus) and a flow-quench technique that has a millisecond time resolution were used to measure the receptor-controlled ion translocation. Using the value of J and the molar concentrations of receptor sites and inorganic ions, we calculated that 6 X 10(3) ions are translocated per msec per receptor. Analysis of electrical noise in frog muscle cells at temperatures above 8 degrees C [Nether, E. & Stevens, C. F. (1977) Annu. Rev. Biophys. Bioeng. 6, 345-381] gave a value of about 1 X 10(4) ions msec-1 per channel. Thus, each technique gives essentially the same result. It is now possible, therefore, to correlate the results obtained when receptor function is measured in two different ways in membrane vesicles and in muscle cells: (i) chemical kinetic measurements, using membrane vesicles, which relate the ligand binding and ion translocation processes and (ii) analysis of acetylcholine noise in muscle cells [Katz, B. & Miledi, R. (1972) J. Physiol. (London) 224, 665-699], which allows one to measure elementary steps in the formation of ion channels through the cell membrane.
已测定乙酰胆碱受体控制的离子转运的比反应速率(J)。在1℃的鳗鱼林格氏液(pH 7.0)中,J = 3×10⁷ M⁻¹ sec⁻¹。J是一个内在常数,是受体的特征,与含受体细胞的其他特性无关,而这些特性也决定离子转运速率。使用膜囊泡(由电鳗的电器官制备)和具有毫秒时间分辨率的流动猝灭技术来测量受体控制的离子转运。利用J的值以及受体位点和无机离子的摩尔浓度,我们计算出每个受体每毫秒转运6×10³个离子。对温度高于8℃的青蛙肌肉细胞中的电噪声进行分析[内瑟,E. & 史蒂文斯,C. F.(1977年)《生物物理与生物工程年度评论》6,345 - 381]得出每个通道约1×10⁴个离子毫秒⁻¹的值。因此,每种技术给出的结果基本相同。所以,现在有可能将在膜囊泡和肌肉细胞中以两种不同方式测量受体功能时获得的结果关联起来:(i)使用膜囊泡进行化学动力学测量,该测量将配体结合和离子转运过程联系起来;(ii)分析肌肉细胞中的乙酰胆碱噪声[卡茨,B. & 米莱迪,R.(1972年)《生理学杂志》(伦敦)224,665 - 699],这使得人们能够测量通过细胞膜形成离子通道的基本步骤。