Botany Department, University College London, Gower St., London WC1, England.
Plant Physiol. 1969 Jan;44(1):60-8. doi: 10.1104/pp.44.1.60.
Optimum conditions for the extraction and assay by ATP-pyrophosphate exchange of the aminoacyl-sRNA synthetases of the various tissues of french bean (Phaseolus vulgaris) seeds and seedlings are described. Extracts of plumules, after passage through Sephadex G-25, were assayed for synthetase activity using an amino acid mixture as substrate, when a 30 to 100-fold stimulation of exchange above the endogenous level was obtained. This marked enhancement of exchange by added amino acids is largely attributed to the use of dilute extracts.The total synthetase activity of the cotyledons was studied in relation to germination. Synthetase activity was highest in the dry cotyledon; activity decreased in proportion to the loss of protein during germination, i.e. the specific activity of the synthetases remained constant for at least the first 6 days of germination. With a few minor exceptions, the specific activity of the individual synthetases remained constant during germination.The absolute activity of the synthetases of the plumule and radicle increased exponentially during germination. The specific activity of the synthetases in both tissues increased approximately 2-fold during the first 2 to 3 days of germination, then gradually decreased. However the asparaginyl-, valyl-, and histidinyl-sRNA synthetases in the plumule did not conform with this general pattern.
描述了从法国菜豆(Phaseolus vulgaris)种子和幼苗的各种组织中提取和通过 ATP-焦磷酸交换测定氨酰-sRNA 合成酶的最佳条件。在通过 Sephadex G-25 后,使用氨基酸混合物作为底物测定新芽提取物中的合成酶活性,此时交换水平比内源性水平高 30 到 100 倍。这种由添加的氨基酸引起的交换的显著增强在很大程度上归因于使用稀释的提取物。与发芽有关,研究了子叶的总合成酶活性。在干子叶中合成酶活性最高;在发芽过程中,随着蛋白质的损失,活性按比例下降,即合成酶的比活性至少在发芽的前 6 天保持恒定。除了一些较小的例外,在发芽过程中,单个合成酶的比活性保持不变。在发芽过程中,芽和根的合成酶的绝对活性呈指数增长。在发芽的前 2 到 3 天,这两种组织中的合成酶的比活性增加了约 2 倍,然后逐渐降低。然而,芽中的天冬酰胺酰基、缬氨酸酰基和组氨酸酰基-sRNA 合成酶不符合这种一般模式。