Sensitive and selective detection of nitric oxide using an H-NOX domain
Sensitive and selective detection of nitric oxide using an H-NOX domain
复制标题
DOI：
10.1021/ja0632714
复制
发表时间：
2006-08-09
影响因子：
15
通讯作者：
Marletta, Michael A.
中科院分区：
文献类型：
--
作者：
Boon, Elizabeth M.;Marletta, Michael A.
TtY140F 是来自腾康热厌氧杆菌的 H−NOX（血红素一氧化氮或氧结合域）蛋白的突变体，是一种新型基因编码的 NO 传感器。TtY140F 很容易纯化，并且可以从标准大肠杆菌中高产率获得。大肠杆菌表达系统。作为未连接的亚铁络合物和亚铁-亚硝酰络合物，它在空气中在一定温度范围内（高达 70 °C）都非常稳定，不会氧化、变性或结合 O2。 NO 结合是定量的，可以使用简单的吸收光谱进行跟踪。理论上，可以准确、轻松地检测 300 nM 至 30 μM 之间的 NO 浓度。
TtY140F, a mutant of the H−NOX (Heme-Nitric Oxide orOXygen binding domain) protein fromThermoanaerobacter tengcongensis, is presented as a novel genetically encoded NO sensor.TtY140F is easily purified and obtained in high yields from standardE. coliexpression systems. It is extremely stable as both the ferrous unligated and ferrous-nitrosyl complexes in air over a range of temperatures (up to 70 °C) without oxidizing, denaturing, or binding O2. NO binding is quantitative and can be followed using simple absorbance spectroscopy. In theory, NO concentrations between 300 nM and 30 μM can be accurately and easily detected.