綠竹筍不同生長時期non-phosphorylating glyceraldehyde- 3-phosphate dehydrogenase之純化與生化性質探討

Abstract

高等植物細胞質中Non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPN) 是一個具有高序列保守性蛋白質，負責催化重要能量及還原物質生成反應。在光合細胞中，此酵素參與了將葉綠體內NADPH運送至細胞質中的機制。為了清楚GAPN在綠竹筍快速生長中所扮演的角色，我們利用蛋白質純化方法，將綠竹筍中GAPN分離出來，並製備專一性的單株抗體。藉由二維電泳及免疫染色方法，我們發現綠竹筍在快速生長過程中，會表現出兩種不同的GAPN (GAPN-A與GAPN-B)，這兩種GAPN存在於在不同的組織中，並具有不同的pI值。這些結果暗示著GAPN-A與GAPN-B在竹筍的快速生長過程中扮演著全然不同的角色。

英文摘要 Non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPN) is a conserved cytosolic protein and catalyze key steps in energy and reducing power partitioning in cells of higher plants. In photosynthetic cells, the enzyme is involved in a shuttle transfer mechanism to export NADPH from the chloroplast to the cytosol. To investigate the role of this enzyme in the rapid-growing green bamboo shoots, we purified and characterized GAPN from green bamboo shoots, and the GAPN specific monoclonal antibody was prepared by using purified GAPN as the antigen. We were identified two kinds of GAPN (GAPN-A and GAPN-B) in the rapid-growing green bamboo shoots by 2-DE and immunostaining. The two kinds of GAPN were found in different tissues during the rapid growth of green bamboo shoots and with distinct pI value. The results suggest that the GAPN-A and GAPN-B may have separate roles in the rapid-growing green bamboo shoots.