Slow-binding inhibition of peptide deformylase by cyclic peptidomimetics as revealed by a new spectrophotometric assay
作者:Kiet T. Nguyen、Xubo Hu、Dehua Pei
DOI:10.1016/j.bioorg.2004.01.001
日期:2004.6
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。