Glitsch H G, Schwarz W, Tappe A, Tierney R, Wilson S, Young L
Arbeitsgruppe Muskelphysiologie, Ruhr-Universitat, Bochum, Germany.
Biochim Biophys Acta. 1996 Jan 31;1278(2):137-46. doi: 10.1016/0005-2736(95)00223-5.
Thermodynamic considerations predict changes of the Na+ pump current (Ip)-voltage (V) relationship of animal cells upon variations of the electrochemical gradients against which cations must be pumped. Experimental data in support of the predictions are sparse. Therefore, the effect on the Ip-V relationship of various electrochemical gradients for pumped Na+ and Cs+ was studied at constant deltaGATP (approximately -39kJ/mol in cardioballs from sheep Purkinje fibres. Control of the subsarcolemmal ionic concentrations during whole-cell recording was ensured by activation of Ip below its half maximal activity or by measuring the initial Ip following reactivation of the Na+/K+ pump. With gradients close to physiological conditions Ip was outward over the entire voltage range and the Ip-V relationship showed a maximum near zero potential. Steepening the ionic gradients diminished the Ip amplitude and outward pump current was no longer detectable between -65 mV and -110 mV. Flattened ionic gradients increased the Ip amplitude and shifted apparently the reversal potential Erev to more negative values. These changes are in line with theoretical considerations. The measured Ip-V relationships were fitted by curves computed on the basis of a simplified Post-Albers scheme of Na+/Cs+ pumping. The increased Ip amplitude at flat ionic gradients was due to a decrease of [Cs+]o for half maximal Ip activation. The maximal Ip amplitude remained unaffected
热力学考量预测,在阳离子必须逆其电化学梯度进行泵转运时,动物细胞的钠泵电流(Ip)-电压(V)关系会发生变化。支持这些预测的实验数据稀少。因此,在恒定的ΔGATP(绵羊浦肯野纤维心球细胞中约为-39kJ/mol)条件下,研究了各种泵转运钠和铯的电化学梯度对Ip-V关系的影响。在全细胞记录过程中,通过将Ip激活至其半数最大活性以下或测量钠/钾泵重新激活后的初始Ip,确保对肌膜下离子浓度的控制。在接近生理条件的梯度下,Ip在整个电压范围内均为外向电流,且Ip-V关系在零电位附近出现最大值。离子梯度变陡会减小Ip幅度,在-65mV至-110mV之间不再能检测到外向泵电流。离子梯度变平缓会增加Ip幅度,并使反转电位Erev明显向更负值偏移。这些变化与理论考量相符。所测得的Ip-V关系通过基于简化的钠/铯泵转运Post-Albers方案计算出的曲线进行拟合。离子梯度平缓时Ip幅度增加是由于半数最大Ip激活时[Cs+]o降低。最大Ip幅度保持不变