Martin-Luther-Universität Halle-Wittenberg, Fachbereich Biochemie/Biotechnologie, Institut für Biochemie, Pflanzenbiochemie, Weinbergweg 16a, Halle, O-4050, Germany.
Plant Physiol. 1992 Apr;98(4):1472-8. doi: 10.1104/pp.98.4.1472.
Four intracellular RNases were found to be induced in cultured tomato (Lycopersicon esculentum) cells upon phosphate starvation. Localization studies revealed three (RNases LV 1-3) in the vacuoles and one (RNase LX) outside these organelles. All of these RNases were purified to homogeneity and were shown to be type I RNases on the basis of type of splitting, substrate, and base specificity at the cleavage site, molecular weight, isoelectric point, and pH optimum. Moreover, RNase LV 3 was shown by fingerprinting of tryptic digests on reversed-phase high-performance liquid chromatography and sequencing the N terminus and two tryptic peptides to be structurally very similar to a recently characterized extracellular RNase LE which is also phosphate regulated (Nürnberger et al. [1990] Plant Physiol 92: 970-976; Jost et al. [1991] Eur J Biochem 198: 1-6). Expression of the four intracellular RNases is induced by depleting the cells of phosphate and repressed by adding phosphate. Our studies indicate that higher plants, in addition to secreting enzymes for scavanging phosphate under starvation conditions, also induce intracellularly emergency rescue systems.
在磷酸盐饥饿的培养番茄(Lycopersicon esculentum)细胞中发现了四种细胞内 RNase。定位研究表明,其中三种(RNase LV1-3)位于液泡中,一种(RNase LX)位于这些细胞器之外。所有这些 RNase 都被纯化到均一性,并根据切割类型、底物和切割位点的碱基特异性、分子量、等电点和 pH 最适值,被证明是 I 型 RNase。此外,通过反向高效液相色谱上的胰蛋白酶消化物指纹图谱分析和 N 末端和两个胰蛋白酶肽的测序,RNase LV3 被证明在结构上与最近表征的也是磷酸盐调节的细胞外 RNase LE 非常相似(Nürnberger 等人,[1990]植物生理学 92:970-976;Jost 等人,[1991]欧洲生物化学杂志 198:1-6)。四种细胞内 RNase 的表达是通过耗尽细胞中的磷酸盐诱导的,而添加磷酸盐则抑制其表达。我们的研究表明,高等植物除了在饥饿条件下分泌用于磷 scavange 的酶外,还诱导细胞内的紧急救援系统。