Braun Y, Hassidim M, Lerner H R, Reinhold L
Department of Botany, The Hebrew University of Jerusalem, Jerusalem, 91904, Israel.
Plant Physiol. 1986 Aug;81(4):1050-6. doi: 10.1104/pp.81.4.1050.
Membrane vesicles were isolated from the roots of the halophyte Atriplex nummularia Lindl. H(+)-translocating Mg(2+)-ATPase activity was manifested by the establishment of a positive membrane potential (measured as SCN(-) accumulation); and also by the establishment of a transmembrane pH gradient (measured by quinacrine fluorescence quenching). H(+)-translocation was highly specific to ATP and was stable to oligomycin. Growing the plants in the presence of 400 millimolar NaCl doubled the proton-translocating activity per milligram of membrane protein and otherwise modulated it in the following ways. First, the flat pH profile observed in non-salt-grown plants was transformed to one showing a peak at about pH 6.2. Second, the lag effect observed at low ATP concentration in curves relating SCN(-) accumulation to ATP concentration was abolished; the concave curvature shown in the double reciprocal plot was diminished. Third, sensitivity to K-2 (N-morpholino)ethanesulfonic acid stimulation was shown in salt-grown plants (about 40% stimulation) but was absent in non-salt-grown plants. Fourth, the KCl concentration bringing about 50% dissipation of ATP-dependent SCN(-) accumulation was 20 millimolar for salt-grown plants and 50 millimolar for non-salt-grown plants. Vanadate sensitivity was shown in both cases. No clear NO(3) (-) inhibition was observed.
从盐生植物滨藜(Atriplex nummularia Lindl.)的根中分离出膜泡。H⁺转运Mg²⁺ -ATP酶活性通过建立正膜电位(以SCN⁻积累来测量)以及建立跨膜pH梯度(通过喹吖因荧光猝灭来测量)得以体现。H⁺转运对ATP具有高度特异性,并且对寡霉素稳定。在400毫摩尔氯化钠存在的情况下培养植物,每毫克膜蛋白的质子转运活性增加了一倍,并且在其他方面以如下方式进行调节。首先,在未用盐培养的植物中观察到的平坦pH曲线转变为在约pH 6.2处出现峰值的曲线。其次,在将SCN⁻积累与ATP浓度相关的曲线中,在低ATP浓度下观察到的滞后效应被消除;双倒数图中显示的凹曲率减小。第三,在盐培养的植物中显示出对K - 2(N - 吗啉代)乙磺酸刺激的敏感性(约40%的刺激),而在未用盐培养的植物中则不存在。第四,使依赖ATP的SCN⁻积累耗散50%的KCl浓度,对于盐培养的植物为20毫摩尔,对于未用盐培养的植物为50毫摩尔。两种情况下均显示出对钒酸盐的敏感性。未观察到明显的NO₃⁻抑制作用。