Zhang Han-Zhong, Kasibhatla Shailaja, Guastella John, Tseng Ben, Drewe John, Cai Sui Xiong
Maxim Pharmaceuticals, 6650 Nancy Ridge Drive, San Diego, California 92121, USA.
Bioconjug Chem. 2003 Mar-Apr;14(2):458-63. doi: 10.1021/bc0256188.
N-Pentafluorobenzoyl-R110 (1a) and N-(2,3,4,5-tetrafluorobenzoyl)-R110 (1b) with enhanced cell retention properties, were prepared from rhodamine 110 (R-110) and the corresponding polyfluorobenzoyl chloride. N-Ac-DEVD-N'-pentafluorobenzoyl-R110 (3a) and N-Ac-DEVD-N'-(2,3,4,5-tetrafluorobenzoyl)-R110 (3b) were prepared as tetrapeptide substrates for caspases. Substrate 3b was efficiently cleaved by human recombinant caspase-3 in an enzyme assay. Substrate 3b also was efficiently cleaved in cell-based apoptosis assays. After cleavage in apoptotic cells by activated caspases, the substrate becomes fluorescent as measured by flow cytometry. These substrates should prove useful in cell-based assays for studying apoptosis inducers and inhibitors.
具有增强细胞保留特性的N-五氟苯甲酰基-R110（1a）和N-（2,3,4,5-四氟苯甲酰基）-R110（1b）由罗丹明110（R-110）和相应的多氟苯甲酰氯制备而成。N-乙酰基-DEVD-N'-五氟苯甲酰基-R110（3a）和N-乙酰基-DEVD-N'-（2,3,4,5-四氟苯甲酰基）-R110（3b）被制备为半胱天冬酶的四肽底物。在酶分析中，底物3b被人重组半胱天冬酶-3有效切割。在基于细胞的凋亡分析中，底物3b也被有效切割。在凋亡细胞中被活化的半胱天冬酶切割后，通过流式细胞术测量，底物会发出荧光。这些底物在用于研究凋亡诱导剂和抑制剂的基于细胞的分析中应该会很有用。
