An Escherichia coli expression-based approach for porphyrin substitution in heme proteins.
An Escherichia coli expression-based approach for porphyrin substitution in heme proteins.
复制标题
一种基于大肠杆菌表达的血红素蛋白中卟啉取代方法。
DOI：
10.1007/978-1-62703-321-3_8
复制
发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Marletta,MichaelA
中科院分区：
文献类型：
--
作者：
Winter,MichaelB;Woodward,JoshuaJ;Marletta,MichaelA
The ability to replace the native heme cofactor of proteins with an unnatural porphyrin of interest affords new opportunities to study heme protein chemistry and engineer heme proteins for new functions. Previous methods for porphyrin substitution rely on removal of the native heme followed by porphyrin reconstitution. However, conditions required to remove the native heme often lead to denaturation, limiting success at heme replacement. An expression-based strategy for porphyrin substitution was developed to circumvent the heme removal and reconstitution steps, whereby unnatural porphyrin incorporation occurs under biological conditions. The approach uses the RP523 strain ofEscherichia coli, which has a deletion of a key gene involved in heme biosynthesis and is permeable to porphyrins. The expression-based strategy for porphyrin substitution detailed here is a robust platform to generate heme proteins containing unnatural porphyrins for diverse applications.
