UV-Vis spectrophotometer or a fluorimeter in a continuous fashion. The utility of the assay was demonstrated by determining the catalytic activity of PDF and the inhibition constants of PDF inhibitors. These studies revealed the slow-binding behavior of a previously reported macrocyclic PDF inhibitor. This method offers several advantages over the existing PDF assays and should be particularly useful
通过将PDF反应与
二肽基肽酶I(
DPPI)偶联并使用N-甲酰基-Met-Lys-
AMC作为底物,开发了一种新的分光光度法/荧光法测定肽去甲酰基酶(PDF)。通过PDF去除N-末端甲酰基使二肽成为
DPPI的有效底物,其随后去除二肽基单元以释放7-
氨基-4-甲基
香豆素作为生色团/荧光团。PDF反应可在UV-Vis分光光度计或荧光计上以连续方式方便地进行监控。通过测定PDF的催化活性和PDF
抑制剂的抑制常数证明了该测定法的实用性。这些研究揭示了以前报道的大环PDF
抑制剂的慢结合行为。