Malhotra B., Glass ADM.
Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z4.
Plant Physiol. 1995 Aug;108(4):1527-1536. doi: 10.1104/pp.108.4.1527.
Potassium influx and cellular [K+] were measured in the unicellular green alga Chlamydomonas reinhardtii after pretreatment in either 10 or 0 mM external K+ ([K]0). K+ (42K+ or 86Rb+) influx was mediated by a saturable, high-affinity transport system (HATS) at low [K+]0 and a linear, low-affinity transport system at high [K+]o. The HATS was typically more sensitive to metabolic inhibition (and darkness) than the low-affinity transport system. Membrane electrical potentials were determined by measuring the equilibrium distribution of tetraphenylphosphonium. These values, together with estimates of cytoplasmic [K+] (B. Malhotra and A.D.M. Glass [1995] Plant Physiol 108: 1537-1545), demonstrated that at 0.1 mM [K+]0 K+ uptake must be active. At higher [K+]0 (>0.3 mM) K+ influx appeared to be passive and possibly channel mediated. When cells were deprived of K+ for 24 h, the Vmax for the HATS increased from 50 x 10-6 to 85 x 10-6 nmol h-1 cell-1 and the Km value decreased from 0.25 to 0.162 mM. Meanwhile, cellular [K+] declined from 24 x 10-6 to 9 x 10-6 nmol cell-1. During this period influx increased exponentially, reaching its peak value after 18 h of K+ deprivation. This increase of K+ influx was not expressed when cells were exposed to inhibitors of protein synthesis. The use of 42K+ and 86Rb+ in parallel experiments demonstrated that Chlamydomonas discriminated in favor of K+ over Rb+, and this effect increased with the duration of K+ deprivation.
在单细胞绿藻莱茵衣藻中,于10 mM或0 mM外部钾离子浓度([K]0)下进行预处理后,测量钾离子流入量和细胞内[K+]。在低[K+]0时,K+(42K+或86Rb+)流入由一个可饱和的高亲和力转运系统(HATS)介导,而在高[K+]o时由一个线性的低亲和力转运系统介导。与低亲和力转运系统相比,HATS通常对代谢抑制(和黑暗)更敏感。通过测量四苯基鏻的平衡分布来确定膜电位。这些值与细胞质[K+]的估计值(B. 马尔霍特拉和A.D.M. 格拉斯[1995]《植物生理学》108: 1537 - 1545)一起表明,在0.1 mM [K+]0时,K+摄取必定是主动的。在较高的[K+]0(>0.3 mM)时,K+流入似乎是被动的,可能由通道介导。当细胞缺钾24小时后,HATS的Vmax从50×10 - 6增加到85×10 - 6 nmol h-1 cell-1,Km值从0.25降至0.162 mM。同时,细胞内[K+]从24×10 - 6降至9×10 - 6 nmol cell-1。在此期间,流入量呈指数增加,在缺钾18小时后达到峰值。当细胞暴露于蛋白质合成抑制剂时,这种K+流入的增加并未表现出来。在平行实验中使用42K+和86Rb+表明,莱茵衣藻对K+的偏好超过Rb+,并且这种效应随着缺钾持续时间的增加而增强。