Urry D W, Trapane T L, Venkatachalam C M
J Membr Biol. 1986;89(1):107-11. doi: 10.1007/BF01870900.
A potassium-39 NMR study of potassium ion interaction with the gramicidin transmembrane channel in phospholipid bilayers at high ion activity is reported which allows determination of a weak binding constant, Kwb approximately equal to 8.3/M, and an off-rate constant for the weak site, kwoff approximately equal to 2.6 X 10(7)/sec. These values are interpreted with the aid of additional NMR data as the binding constant for formation of the doubly occupied channel state and the rate constant for an ion leaving the doubly occupied state. Considering the singly occupied channel state for the potassium ion to be "electrically silent" at 1 molar ion activity, as with the sodium ion, the single-channel conductance for 100 mV and 30 degrees C calculated to be 29 pS, and using the same approximation with previous NMR results on the sodium and rubidium ions, reasonable conductance ratios were calculated. Further experimental estimates of the other three constants with the experimental location of binding sites and Eyring rate theory to introduce voltage dependence allowed a more complete calculation of the two-site channel. The single-channel conductance for potassium ion is calculated to be 24 pS at 1 M activity and 26 pS at 0.6 M activity, which compares for diphytanoyl phosphatidylcholine membranes to an experimental most probable single-channel conductance of 25 pS and a mean channel conductance of 20 pS. The calculated conductance ratios using NMR-derived constants were gamma (K)/gamma (Na) = 2.0 and gamma (Rb)/gamma (Na) = 4.3.(ABSTRACT TRUNCATED AT 250 WORDS)
本文报道了在高离子活性下,磷脂双层中钾离子与短杆菌肽跨膜通道相互作用的钾-39核磁共振研究。该研究确定了一个弱结合常数Kwb约为8.3/M,以及弱结合位点的解离速率常数kwoff约为2.6×10⁷/秒。借助其他核磁共振数据,这些值被解释为形成双占据通道状态的结合常数以及离子离开双占据状态的速率常数。考虑到在1摩尔离子活性下,钾离子的单占据通道状态如同钠离子一样“电沉默”,计算出在100 mV和30℃时单通道电导为29 pS,并采用与先前关于钠离子和铷离子的核磁共振结果相同的近似方法,计算出了合理的电导比。通过结合位点的实验位置以及艾林速率理论对其他三个常数进行进一步的实验估计,引入电压依赖性后,对双位点通道进行了更完整的计算。计算得出在1 M活性下钾离子的单通道电导为24 pS,在0.6 M活性下为26 pS,对于二植烷酰磷脂酰胆碱膜,实验测得的最可能单通道电导为25 pS,平均通道电导为20 pS。使用核磁共振衍生常数计算出的电导比为γ(K)/γ(Na)=2.0,γ(Rb)/γ(Na)=4.3。(摘要截短于250字)