| Abstract: | Basic nuclear magnetic resonance (NMR) features of 23Na ions bound to the gramicidin channel (packaged into lecithin liposomes) were studied. The first binding constant K1 of Na+ was not significantly dependent on channel models employed. With the two-identical-site model (Model I), K1 was 13.7 (+/- 1.4) molal-1 (in the activity basis) at 25 degrees C; when the binding of a third ion was included (Model II), it was 13.0 (+/- 2.0) molal-1. The second binding constant K2 was model dependent; it was 1.6 (+/- 0.2) and 3-4 molal-1 for Models I and II, respectively. The rate constants, k-1 and k-2, of Na+ for exit from singly and doubly loaded channels, respectively, were 8 X 10(5) s-1 less than or equal to k-1 less than or equal to 3 X 10(6) s-1 and 8 X 10(5) s-1 less than or equal to k-2 less than or equal to 1.0 X 10(7) s-1 at 25 degrees C; the lower bound represents a rough approximation of k-1. The ratio k-2/k-1 was greater than one and did not greatly exceed 20. From the competition experiment, K1 of T1+ was 5.7 (+/- 0.6) X 10(2) molal-1. The longitudinal relaxation time T1 of bound 23Na in the state of single occupancy (T 1B sing) was virtually independent of models, 0.56 (+/- 0.03) and 0.55 (+/- 0.04) ms at 25 degrees C for Models I and II, respectively. For the state of double occupancy, T1 of bound 23Na (T 1B doub) was model dependent: 0.27 (+/- 0.01) and 0.4-0.6 ms for Models I and II. The correlation time tau c of bound 23Na was 2.2 (+/- 0.2) ns at 25 degrees C for single occupancy; tau c for double occupancy was not significantly different from this value. The estimated tau c was found to involve no appreciable contribution of the exchange of 23Na between the channel and the bulk solution. Thé quadrupole coupling constant chi was 1.0 (+/- 0.1) MHz for 23Na in single occupancy; chi for double occupancy was 0.9-1.4 MHz, depending on models. A lower bound of the average quadrupole coupling constant chi alpha was 0.13-0.26 MHz at 25 degrees C for 23Na in single occupancy; this value represents a rough approximation of chi alpha at this temperature. An argument based on the estimated chi alpha and the known conformation of the gramicidin channel suggests that the binding site is a small domain near the channel end.(ABSTRACT TRUNCATED AT 400 WORDS) |
