Graduate School of Life Science and Systems Engineering, Kyushu Institute of Technology, 2-4 Hibikino, Wakamatsu-ku, Kitakyushu, Fukuoka 808-0196, Japan.
Graduate School of Life Science and Systems Engineering, Kyushu Institute of Technology, 2-4 Hibikino, Wakamatsu-ku, Kitakyushu, Fukuoka 808-0196, Japan.
Bioorg Med Chem. 2018 Jul 23;26(12):3468-3473. doi: 10.1016/j.bmc.2018.05.021. Epub 2018 May 18.
Peptide substrates were double labeled with pyrenes to prepare fluorescent probes for highly sensitive detection of protease activity and evaluation of protease inhibitors using pyrene monomer/excimer signals. Two proximate pyrene moieties formed excited state dimers in the probes, and these pyrene excimer formations were dissociated by tryptic digestion. The specificity constant of the optimum bispyrene peptide probe was 2.7 times higher than that of the conventional peptide-4-methylcoumarin amide. Moreover, our probe had high sensitivity with an estimated detection limit for trypsin of 4.11 pM. The half maximal inhibitory concentration and dissociation constant of the Bowman-Birk inhibitor were successfully estimated.
肽底物用芘双重标记，制备荧光探针，用于高灵敏度检测蛋白酶活性，并利用芘单体/激基复合物信号评估蛋白酶抑制剂。两个邻近的芘基团在探针中形成激发态二聚体，这些芘激基复合物的形成通过胰蛋白酶消化而解离。最佳双芘肽探针的特异性常数比传统肽-4-甲基香豆素酰胺高 2.7 倍。此外，我们的探针具有高灵敏度，估计对胰蛋白酶的检测限为 4.11 pM。成功评估了 Bowman-Birk 抑制剂的半最大抑制浓度和解离常数。
