Yoshioka K, Saito M, Oh K B, Nemoto Y, Matsuoka H, Natsume M, Abe H
Department of Biotechnology, Faculty of Technology, Tokyo University of Agriculture and Technology, Japan.
Biosci Biotechnol Biochem. 1996 Nov;60(11):1899-901. doi: 10.1271/bbb.60.1899.
A fluorescent derivative of D-glucose, 2-NBDG, which was previously developed for the evaluation of glucose uptake activity by living cells, was used on Escherichia coli cells and its fate after incorporation in the cells was investigated. 2-NBDG was converted to another fluorescent derivative (2-NBDG metabolite) immediately after it was taken by E. coli cells. This 2-NBDG metabolite was then decomposed to non-fluorescent forms. 2-NBDG metabolite was decomposed into the original 2-NBDG by G6Pase with concurrent liberation of inorganic phosphate. Furthermore, FAB/MS analysis showed that its molecular weight was 420, the same value as that of 2-NBDG 6-phosphate. These indicate 2-NBDG metabolite should be 2-NBDG 6-phosphate. Based on these results, the feasibility of 2-NBDG as a fluorescent non-toxic probe for glucose uptake activity and its application to viability assessment of various living systems are discussed.
一种D - 葡萄糖的荧光衍生物2 - NBDG，其先前被开发用于评估活细胞的葡萄糖摄取活性，被应用于大肠杆菌细胞，并研究了其掺入细胞后的命运。2 - NBDG被大肠杆菌细胞摄取后立即转化为另一种荧光衍生物（2 - NBDG代谢物）。然后这种2 - NBDG代谢物分解为非荧光形式。2 - NBDG代谢物被葡萄糖 - 6 - 磷酸酶分解为原始的2 - NBDG，同时释放出无机磷酸盐。此外，快原子轰击质谱（FAB/MS）分析表明其分子量为420，与6 - 磷酸 - 2 - NBDG的分子量相同。这些表明2 - NBDG代谢物应该是6 - 磷酸 - 2 - NBDG。基于这些结果，讨论了2 - NBDG作为葡萄糖摄取活性的荧光无毒探针的可行性及其在各种生物系统活力评估中的应用。
