Pouliquin P, Boyer J C, Grouzis J P, Gibrat R
Biochimie et Physiologie Mol¿eculaire des Plantes, Agro-M/CNRS (Unit¿e Mixte de Recherche 5004)/Institut National de la Recherche Agronomique/Universit¿e de Montpellier 11, 2, Place Viala, F-34060 Montpellier, France.
Plant Physiol. 2000 Jan;122(1):265-74. doi: 10.1104/pp.122.1.265.
The net initial passive flux (J(Ni)) in reconstituted plasma membrane (PM) vesicles from maize (Zea mays) root cells was measured as recently described (P. Pouliquin, J.-P. Grouzis, R. Gibrat ¿1999 Biophys J 76: 360-373). J(Ni) in control liposomes responded to membrane potential or to NO(3)(-) as expected from the Goldman-Hodgkin-Katz diffusion theory. J(Ni) in reconstituted PM vesicles exhibited an additional component (J(Nif)), which was saturable (K(m) for NO(3)(-) approximately 3 mM, with J(Nifmax) corresponding to 60 x 10(-9) mol m(-2) s(-1) at the native PM level) and selective (NO(3)(-) = ClO(3)(-) > Br(-) > Cl(-) = NO(2)(-); relative fluxes at 5 mM: 1:0.34:0.19). J(Nif) was totally inhibited by La(3+) and the arginine reagent phenylglyoxal. J(Nif) was voltage dependent, with an optimum voltage at 105 mV at pH 6.5. The activation energy of J(Nif) was high (129 kJ mol(-1)), close to that of the H(+)-ATPase (155 kJ mol(-1)), and J(Nif) displayed the same acidic optimal pH (pH 6.5) as that of the H(+) pump. This is the first example, to our knowledge, of a secondary transport at the plant PM with such a feature. Several properties of the NO(3)(-) uniport seem poorly compatible with that reported for plant anion channels and to be attributable instead to a classical carrier. The physiological relevance of these findings is suggested.
按照最近描述的方法(P. Pouliquin、J.-P. Grouzis、R. Gibrat,1999年,《生物物理学杂志》76卷:360 - 373页),测量了玉米(Zea mays)根细胞重构质膜(PM)囊泡中的净初始被动通量(J(Ni))。对照脂质体中的J(Ni)对膜电位或NO₃⁻的响应符合戈德曼 - 霍奇金 - Katz扩散理论的预期。重构PM囊泡中的J(Ni)表现出一个额外的成分(J(Nif)),它是可饱和的(NO₃⁻的K(m)约为3 mM,在天然PM水平下J(Nifmax)对应于60×10⁻⁹ mol m⁻² s⁻¹)且具有选择性(NO₃⁻ = ClO₃⁻ > Br⁻ > Cl⁻ = NO₂⁻;5 mM时的相对通量:1:0.34:0.19)。J(Nif)完全被La³⁺和精氨酸试剂苯乙二醛抑制。J(Nif)与电压有关,在pH 6.5时最佳电压为105 mV。J(Nif)的活化能很高(129 kJ mol⁻¹),接近H⁺ - ATP酶的活化能(155 kJ mol⁻¹),并且J(Nif)表现出与H⁺泵相同的酸性最佳pH(pH 6.5)。据我们所知,这是植物PM上具有这种特征的二次转运的首个例子。NO₃⁻单向转运体的几个特性似乎与报道的植物阴离子通道的特性不太相符,而更应归因于一种经典载体。这些发现的生理相关性也得到了提示。