Cortès Sandra, Gromova Marina, Evrard Adeline, Roby Claude, Heyraud Alain, Rolin Dominique B, Raymond Philippe, Brouquisse Renaud M
Commissariat à l'Energie Atomique, Centre National de la Recherche Scientifique, Université Joseph Fourier, Unité Mixte de Recherche 5019 Physiologie Cellulaire Végétale, 17 rue des Martyrs, 38054 Grenoble cedex 9, France.
Plant Physiol. 2003 Feb;131(2):824-37. doi: 10.1104/pp.010538.
In plants, sugars are the main respiratory substrates and important signaling molecules in the regulation of carbon metabolism. Sugar signaling studies suggested that sugar sensing involves several key components, among them hexokinase (HXK). Although the sensing mechanism of HXK is unknown, several experiments support the hypothesis that hexose phosphorylation is a determining factor. Glucose (Glc) analogs transported into cells but not phosphorylated are frequently used to test this hypothesis, among them 3-O-methyl-Glc (3-OMG). The aim of the present work was to investigate the effects and fate of 3-OMG in heterotrophic plant cells. Measurements of respiration rates, protein and metabolite contents, and protease activities and amounts showed that 3-OMG is not a respiratory substrate and does not contribute to biosynthesis. Proteolysis and lipolysis are induced in 3-OMG-fed maize (Zea mays L. cv DEA) roots in the same way as in sugar-starved organs. However, contrary to the generally accepted idea, phosphorous and carbon nuclear magnetic resonance experiments and enzymatic assays prove that 3-OMG is phosphorylated to 3-OMG-6-phosphate, which accumulates in the cells. Insofar as plant HXK is involved in sugar sensing, these findings are discussed on the basis of the kinetic properties because the catalytic efficiency of HXK isolated from maize root tips is five orders of magnitude lower for 3-OMG than for Glc and Man.
在植物中,糖类是主要的呼吸底物,也是调节碳代谢的重要信号分子。糖类信号研究表明,糖感知涉及几个关键组分,其中包括己糖激酶(HXK)。尽管HXK的感知机制尚不清楚,但多项实验支持己糖磷酸化是一个决定性因素这一假说。经常使用转运到细胞中但未被磷酸化的葡萄糖类似物来检验这一假说,其中包括3 - O - 甲基葡萄糖(3 - OMG)。本研究的目的是探究3 - OMG在异养植物细胞中的作用和去向。对呼吸速率、蛋白质和代谢物含量以及蛋白酶活性和数量的测量结果表明,3 - OMG不是呼吸底物,也不参与生物合成。在以3 - OMG饲喂的玉米(Zea mays L. cv DEA)根中,蛋白水解和脂肪分解的诱导方式与在糖饥饿器官中的诱导方式相同。然而,与普遍接受的观点相反,磷和碳核磁共振实验以及酶促分析证明,3 - OMG被磷酸化为3 - OMG - 6 - 磷酸,并在细胞中积累。鉴于植物HXK参与糖感知,基于动力学性质对这些发现进行了讨论,因为从玉米根尖分离的HXK对3 - OMG的催化效率比对葡萄糖和甘露糖的催化效率低五个数量级。