Spalding E P, Slayman C L, Goldsmith M H, Gradmann D, Bertl A
Department of Biology, Kline Biology Tower, Yale University, New Haven, Connecticut 06511.
Plant Physiol. 1992 May;99(1):96-102. doi: 10.1104/pp.99.1.96.
White light (25 watts per square meter) induced an increase in plasma membrane K(+)-channel activity and a 30- to 70-millivolt transient membrane depolarization (completed in 2-3 minutes) in Arabidopsis thaliana leaf mesophyll cells. Transport characteristics of three types of ion channels in the plasma membrane were determined using inside-out patches. With 220 millimolar K(+) on the cytoplasmic side of the patch and 50 millimolar K(+) in the pipette, (220/50 K), the open-channel current-voltage curves of these channels were sigmoidal and consistent with an enzyme kinetic model. Two channel types were selective for K(+) over Na(+) and Cl(-). One (named PKC1) had a maximum conductance (G(max)) of 44 picosiemens at a membrane voltage (V(m)) of -65 mV in (220/50 K) and is stimulated by light. The other (PKC2) had G(max) = 66 picosiemens at V(m) = 60 millivolts in (220/50 K). The third channel type (PCC1) transported K(+) and Na(+) about equally well but not Cl(-). It had G(max) = 109 picosiemens at V(m) = 55 millivolts in (250/50 K) with 10 millimolar Ca(2+) on the cytoplasmic side. Reducing Ca(2+) to 0.1 millimolar increased PCC1 open-channel currents by approximately 50% in a voltage-independent manner. Averaged over time, PKC2 and PCC1 currents strongly outward rectified and PKC1 currents did so weakly. Reductants (1 millimolar dithiothreitol or 10 millimolar beta-mercaptoethanol) added to the cytoplasmic side of an excised patch increased the open probability of all three channel types.
白光(每平方米25瓦)可诱导拟南芥叶肉细胞的质膜钾离子通道活性增加,并使膜发生30至70毫伏的瞬时去极化(2至3分钟内完成)。使用内向外膜片测定质膜中三种离子通道的转运特性。当膜片胞质侧为220毫摩尔钾离子,移液管中为50毫摩尔钾离子(220/50 K)时,这些通道的开放通道电流-电压曲线呈S形,符合酶动力学模型。两种通道类型对钾离子的选择性高于钠离子和氯离子。其中一种(命名为PKC1)在膜电压(V(m))为-65毫伏(220/50 K)时,最大电导(G(max))为44皮西门子,且受光刺激。另一种(PKC2)在V(m) = 60毫伏(220/50 K)时,G(max) = 66皮西门子。第三种通道类型(PCC1)对钾离子和钠离子的转运能力相当,但对氯离子无转运能力。在胞质侧有10毫摩尔钙离子的情况下,当V(m) = 55毫伏(250/50 K)时,其G(max) = 109皮西门子。将钙离子浓度降至0.1毫摩尔可使PCC1的开放通道电流以电压非依赖方式增加约50%。随着时间平均,PKC2和PCC1电流强烈外向整流,而PKC1电流则较弱。添加到切除膜片胞质侧的还原剂(1毫摩尔二硫苏糖醇或10毫摩尔β-巯基乙醇)可增加所有三种通道类型的开放概率。