We report the synthesis of seven new proluciferins for convenient activity determination of enzymes belonging to the cytochrome P450 (CYP) 4 family. Biotransformation of these probe substrates was monitored using each of the twelve human CYP4 family members, and eight were found to act at least on one of them. For all substrates, activity of CYP4Z1 was always highest, while that of CYP4F8 was always second highest. Site of metabolism (SOM) predictions involving SMARTCyp and docking experiments helped to rationalize the observed activity trends linked to substrate accessibility and reactivity. We further report the first homology model of CYP4F8 including suggested substrate recognition residues in a catalytically competent conformation accessed by replica exchange solute tempering (REST) simulations.

我们报告了七个新的proluciferins方便活动测定属于细胞色素P450（CYP）4家族的酶的合成。使用十二种人类CYP4家族成员中的每一个监测这些探针底物的生物转化，发现其中八种至少对其中一种起作用。对于所有底物，CYP4Z1的活性始终最高，而CYP4F8的活性始终第二高。涉及SMARTCyp和对接实验的代谢位点（SOM）预测有助于合理化观察到的与底物可及性和反应性有关的活动趋势。我们进一步报告了CYP4F8的第一个同源性模型，包括通过复制交换溶质回火（REST）模拟访问的催化有效构象中的建议底物识别残基。