Optical detection of cytochrome P450 by sensitizer-linked substrates
Optical detection of cytochrome P450 by sensitizer-linked substrates
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DOI：
10.1073/pnas.96.23.12987
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发表时间：
1999-11-09
影响因子：
11.1
通讯作者：
Gray, HB
中科院分区：
文献类型：
--
作者：
Dmochowski, IJ;Crane, BR;Gray, HB
在复杂介质中检测、表征和操纵特定生物分子的能力对于理解代谢过程至关重要。特别重要的目标是参与药物代谢和许多疾病状态的加氧酶（细胞色素P450），包括肝脏和肾脏功能障碍，神经系统疾病和癌症。我们已经发现，钌光敏剂连接到P450基板特异性识别亚微摩尔细胞色素P450（凸轮）在其他血红素蛋白的存在下。在P450：Ru-底物缀合物中，向血红素的能量转移显著地加速了Ru-发光衰减。P450（cam）：Ru-金刚烷基络合物的晶体结构揭示了通过通道进入活性中心，该通道的深度（Ru-Fe距离为21埃）与从能量转移动力学分析中提取的通道几乎相同。适当构建的敏化剂连接的基板库可以用来探测掩埋的活性位点的空间和电子性质。
The ability to detect, characterize, and manipulate specific biomolecules in complex media is critical for understanding metabolic processes. Particularly important targets are oxygenases (cytochromes P450) involved in drug metabolism and many disease states, including liver and kidney dysfunction, neurological disorders, and cancer. We have found that Ru photosensitizers linked to P450 substrates specifically recognize submicromolar cytochrome P450(cam) in the presence of other heme proteins. In the P450:Ru-substrate conjugates, energy transfer to the heme dramatically accelerates the Ru-luminescence decay, The crystal structure of a P450(cam):Ru-adamantyl complex reveals access to the active center via a channel whose depth (Ru-Fe distance is 21 Angstrom) is virtually the same as that extracted from an analysis of the energy-transfer kinetics. Suitably constructed libraries of sensitizer-linked substrates could be employed to probe the steric and electronic properties of buried active sites.