Specific reaction rate of acetylcholine receptor-controlled ion translocation: a comparison of measurements with membrane vesicles and with muscle cells.
Author(s) -
George P. Hess,
Hitoshi Aoshima,
Derek J. Cash,
Bernard Lenchitz
Publication year - 1981
Publication title -
proceedings of the national academy of sciences
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.78.3.1361
Subject(s) - electrophorus , acetylcholine receptor , vesicle , biophysics , chemistry , acetylcholine , ion , receptor , membrane potential , ion channel , membrane , analytical chemistry (journal) , biochemistry , torpedo , biology , chromatography , organic chemistry , endocrinology
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.
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