The ferrous-oxy complex of human aromatase
The ferrous-oxy complex of human aromatase
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DOI：
10.1016/j.bbrc.2008.05.011
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发表时间：
2008-07-25
影响因子：
3.1
通讯作者：
Sligar, Stephen G.
中科院分区：
文献类型：
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作者：
Grinkova, Yelena V.;Denisov, Ilia G.;Sligar, Stephen G.
在这篇文章中，我们记录了异源表达的截短人芳香酶（CYP 19）自组装成纳米级磷脂双层（Nanodisks）。所得的P450 CYP 19制剂是稳定的，并且可以与底物雄烯二酮紧密结合以形成几乎完整的高自旋铁蛋白。在快速扫描停止流中，将Nanodisks中的亚铁CYP 19与大气分子氧厌氧混合，并观察到亚铁-氧复合物的形成。一阶衰减的氧复合物释放超氧化物和再生的铁酶进行了动力学监测。令人惊讶的是，芳香酶的亚铁-氧复合物比肝脏CYP 3A 4的亚铁-氧复合物更稳定，这为精确确定这一重要人类药物靶标中反应周期中间体的生化和生物物理性质开辟了道路。(C)2008年爱思唯尔公司All rights reserved.
In this communication, we document the self-assembly of heterologously expressed truncated human aromatase (CYP19) into nanometer scale phospholipids bilayers (Nanodiscs). The resulting P450 CYP19 preparation is stable and can tightly associate with the substrate androstenedione to form a nearly complete high-spin ferric protein. Ferrous CYP19 in Nanodiscs was mixed anaerobically in a rapid-scan stopped-flow with atmospheric dioxygen and the formation of the ferrous-oxy complex observed. First order decay of the oxy-complex to release superoxide and regenerate the ferric enzyme was monitored kinetically. Surprisingly, the ferrous-oxy complex of aromatase is more stable than that of hepatic CYP3A4, opening the path to precisely determine the biochemical and biophysical properties of the reaction cycle intermediates in this important human drug target. (C) 2008 Elsevier Inc. All rights reserved.