IMMUNOCYTOCHEMICAL LOCALIZATION OF PHASEOLIN AND PHYTOHEMAGGLUTININ IN THE ENDOPLASMIC-RETICULUM AND GOLGI-COMPLEX OF DEVELOPING BEAN COTYLEDONS
IMMUNOCYTOCHEMICAL LOCALIZATION OF PHASEOLIN AND PHYTOHEMAGGLUTININ IN THE ENDOPLASMIC-RETICULUM AND GOLGI-COMPLEX OF DEVELOPING BEAN COTYLEDONS
复制标题
DOI：
10.1007/bf00402940
复制
发表时间：
1985-01-01
期刊：
影响因子：
4.3
通讯作者：
CHRISPEELS, MJ
中科院分区：
文献类型：
--
作者：
GREENWOOD, JS;CHRISPEELS, MJ
豆科植物种子的发育伴随着贮藏蛋白和凝集素的合成，以及这些蛋白在蛋白质贮藏泡（蛋白体）中的沉积。本文报道了普通菜豆（Phaseolus vulgaris L.）种子发育过程中的亚细胞分布，主要贮藏蛋白（菜豆素）和主要凝集素（植物血凝素，PHA）的含量进行了检测。使用间接免疫细胞化学方法定位蛋白质，其中用PHA或菜豆蛋白特异性的兔抗体对冷冻切片进行免疫标记。然后使用吸附在4至5 nm胶体Au颗粒上的山羊抗兔免疫球蛋白G定位结合的抗体。切片用OsO4后固定，脱水，并包埋在网格上的塑料中。PHA和菜豆蛋白在贮藏薄壁细胞中的分布相似，主要存在于发育中的蛋白体中。内质网和高尔基复合体（池堆栈和相关的囊泡）也被特异性标记为这两种蛋白质，而细胞质和其他细胞器，如线粒体，没有。这些观察结果支持了这样的假设，即贮藏蛋白和凝集素从其合成位点粗面内质网到其沉积位点蛋白体的运输是由高尔基复合体介导的。
Development of legume seeds is accompanied by the synthesis of storage proteins and lectins, and the deposition of these proteins in protein-storage vacuoles (protein bodies). The subcellular distribution, in developing seeds of the common bean, Phaseolus vulgaris L., of the major storage protein (phaseolin) and the major lectin (phytohemagglutinin, PHA) was examined. The proteins were localized using an indirect immunocytochemical method in which ultrathin frozen sections were immunolabeled with rabbit antibodies specific for either PHA or phaseolin. Bound antibodies were then localized using goat-anti-rabbit immunoglobulin G adsorbed onto 4- to 5-nm colloidal Au particles. The sections were post-fixed with OsO4, dehydrated, and embedded in plastic on the grids. Both PHA and phaseolin exhibited a similar distribution in the storage-parenchyma cells, being found primarily in the developing protein bodies. Endoplasmic reticulum and Golgi complexes (cisternal stacks and associated vesicles) also were specifically labeled for both proteins, whereas the cytosol and other organelles, such as mitochondria, were not. These observations support the hypothesis that the transport of storage proteins and lectins from their site of synthesis, the rough endoplasmic reticulum, to their site of deposition, the protein bodies, is mediated by the Golgi complex.