scholarly journals Direct Measurement of K+ Channels in Thylakoid Membranes by Incorporation of Vesicles into Planar Lipid Bilayers

1989 ◽  
Vol 91 (1) ◽  
pp. 249-252 ◽  
Author(s):  
Mark Tester ◽  
Michael R. Blatt
1988 ◽  
Vol 92 (5) ◽  
pp. 549-567 ◽  
Author(s):  
J Neyton ◽  
C Miller

Single high-conductance Ca2+-activated K+ channels from rat skeletal muscle were inserted into planar lipid bilayers, and discrete blocking by the Ba2+ ion was studied. Specifically, the ability of external K+ to reduce the Ba2+ dissociation rate was investigated. In the presence of 150 mM internal K+, 1-5 microM internal Ba2+, and 150 mM external Na+, Ba2+ dissociation is rapid (5 s-1) in external solutions that are kept rigorously K+ free. The addition of external K+ in the low millimolar range reduces the Ba2+ off-rate 20-fold. Other permeant ions, such as Tl+, Rb+, and NH4+ show a similar effect. The half-inhibition constants rise in the order: Tl+ (0.08 mM) less than Rb+ (0.1 mM) less than K+ (0.3 mM) less than Cs+ (0.5 mM) less than NH4+ (3 mM). When external Na+ is replaced by 150 mM N-methyl glucamine, the Ba2+ off-rate is even higher, 20 s-1. External K+ and other permeant ions reduce this rate by approximately 100-fold in the micromolar range of concentrations. Na+ also reduces the Ba2+ off-rate, but at much higher concentrations. The half-inhibition concentrations rise in the order: Rb+ (4 microM) less than K+ (19 microM) much less than Na+ (27 mM) less than Li+ (greater than 50 mM). The results require that the conduction pore of this channel contains at least three sites that may all be occupied simultaneously by conducting ions.


Diabetologia ◽  
1992 ◽  
Vol 35 (7) ◽  
pp. 619-623 ◽  
Author(s):  
Y. Oosawa ◽  
S. J. H. Ashcroft ◽  
F. M. Ashcroft

FEBS Letters ◽  
2005 ◽  
Vol 579 (7) ◽  
pp. 1675-1682 ◽  
Author(s):  
Karen Castillo ◽  
Juan Bacigalupo ◽  
Daniel Wolff

1984 ◽  
Vol 46 (6) ◽  
pp. 831-835 ◽  
Author(s):  
L. Olans ◽  
S. Sariban-Sohraby ◽  
D.J. Benos

2015 ◽  
Vol 30 (3) ◽  
pp. 1306-1316 ◽  
Author(s):  
Kunitoshi Uchida ◽  
Lusine Demirkhanyan ◽  
Swapna Asuthkar ◽  
Alejandro Cohen ◽  
Makoto Tominaga ◽  
...  

1997 ◽  
Vol 273 (2) ◽  
pp. H796-H804 ◽  
Author(s):  
C. Valdivia ◽  
J. O. Hegge ◽  
R. D. Lasley ◽  
H. H. Valdivia ◽  
R. Mentzer

We investigated the effects of myocardial stunning on the function of the two main Ca2+ transport proteins of the sarcoplasmic reticulum (SR), the Ca(2+)-adenosinetriphosphatase and the Ca(2+)-release channel or ryanodine receptor. Regional myocardial stunning was induced in open-chest pigs (n = 6) by a 10-min occlusion of the left anterior descending coronary artery (LAD) and 2 h reperfusion. SR vesicles isolated from the LAD-perfused region (stunned) and the normal left circumflex coronary artery (LC)-perfused region were used to assess the oxalate-supported 45Ca2+ uptake, [3H]ryanodine binding, and single-channel recordings of ryanodine-sensitive Ca(2+)-release channels in planar lipid bilayers. Myocardial stunning decreased LAD systolic wall thickening to 20% of preischemic values. The rate of SR 45Ca2+ uptake in the stunned LAD bed was reduced by 37% compared with that of the normal LC bed (P < 0.05). Stunning was also associated with a 38% reduction in the maximal density of high-affinity [3H]ryanodine binding sites (P < 0.05 vs. normal LC) but had no effect on the dissociation constant. The open probability of ryanodine-sensitive Ca(2+)-release channels determined by single channel recordings in planar lipid bilayers was 26 +/- 2% for control SR (n = 33 channels from 3 animals) and 14 +/- 2% for stunned SR (n = 21 channels; P < 0.05). This depressed activity of SR function observed in postischemic myocardium could be one of the mechanisms underlying myocardial stunning.


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