Abstract：
Using myoglobin (Mb) as a model protein, we herein developed a facial approach to modifying the heme active site. A cavity was first generated in the heme distal site by F46 C mutation, and the thiol group of Cys46 was then used for covalently linked to exogenous ligands, 1H-1,2,4-triazole-3-thiol and 1-(4-hydroxyphenyl)-1H-pyrrole-2,5-dione. The engineered proteins, termed F46C-triazole Mb and F46C-phenol Mb, respectively, were characterized by X-ray crystallography, spectroscopic and stopped-flow kinetic studies. The results showed that both the heme coordination state and the protein function such as H2 O2 activation and peroxidase activity could be efficiently regulated, which suggests that this approach might be generally applied to the design of functional heme proteins.
摘要：
使用肌红蛋白（Mb）作为模型蛋白质，我们在此开发了一种面部方法来修饰血红素活性位点。由F46C突变首先在血红素远端部位产生空腔，然后将Cys46的巯基与外源配体共价连接，1H-1,2,4-三唑-3-硫醇和1-(4-羟基苯基)-1H-吡咯-2,5-二酮。工程蛋白质，称为F46C-三唑Mb和F46C-苯酚Mb，分别,通过X射线晶体学表征，光谱和停流动力学研究。结果表明，血红素的配位状态和蛋白质的功能，如H2O2的活性和过氧化物酶的活性，可以有效地调节，这表明这种方法可能普遍适用于功能性血红素蛋白的设计。
