McLean M A, Maves S A, Weiss K E, Krepich S, Sligar S G
Beckman Institute for Advanced Science and Technology, University of Illinois, Urbana, Illinois, 61801, USA.
Biochem Biophys Res Commun. 1998 Nov 9;252(1):166-72. doi: 10.1006/bbrc.1998.9584.
We report the cloning, expression, purification, and molecular characterization of a cytochrome P450 (CYP119) from the thermophilic archaea Sulfolobus solfataricus. This protein displays an absorption spectra in the reduced, oxidized, and carbonyl adduct analogous to those of other P450 enzymes. We demonstrate that P450 (CYP119) exhibits remarkable thermo- and pressure stability, with a melting temperature 40 degrees higher than that of the extensively studied cytochrome P450cam (CYP101) and an optical spectra completely resistant to the formation of the inactive P420 by hydrostatic pressure up to 2 kbar. CO flash photolysis experiments, as well as construction of a CYP119 homology model, suggest an open active site with greater solvent access than P450 (CYP101) and similar to that of P450 (CYP102). This communication represents the first molecular characterization of an extremophilic cytochrome P450.
我们报道了来自嗜热古菌嗜热栖热菌（Sulfolobus solfataricus）的一种细胞色素P450（CYP119）的克隆、表达、纯化及分子特征。该蛋白在还原态、氧化态和羰基加合物状态下的吸收光谱与其他P450酶类似。我们证明P450（CYP119）表现出显著的热稳定性和压力稳定性，其解链温度比经过广泛研究的细胞色素P450cam（CYP101）高40摄氏度，并且其光谱在高达2千巴的静水压力下完全不会形成无活性的P420。CO快速光解实验以及CYP119同源模型的构建表明，其活性位点开放，与P450（CYP101）相比溶剂可及性更高，与P450（CYP102）类似。本通讯报道了嗜极端细胞色素P450的首次分子特征。
