期刊
JOURNAL OF INORGANIC BIOCHEMISTRY
卷 104, 期 10, 页码 1071-1078出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.jinorgbio.2010.06.004
关键词
Azurin; Blue copper; Type 1 copper; Reduction potential; Secondary coordination sphere; Electrochemistry
资金
- Swenson Family Foundation
- UM-GIA
- Camille and Henry Dreyfus Foundation
Recent evidence has shown that the properties of metal binding sites can be tuned by more than the ligands in the primary coordination sphere. We investigated the incorporation of four phenylalanine residues into the secondary coordination sphere of the small soluble blue copper protein azurin. The locations for placement of these residues in azurin were based on the structure of the highly hydrophobic blue copper protein rusticyanin, which is known to have a significantly higher reduction potential than azurin. Using site-directed mutagenesis, these residues in close proximity to the copper binding site were mutated to large hydrophobic phenylalanine residues individually and in combination. We also added the Met121Leu mutation on top of the Phe mutations to construct a total of 13 variants. We found little change in the UV-visible absorption and EPR data for these proteins, however modest increases in reduction potential were observed with increases by as much as 30 mV per Phe residue. Furthermore, we observed the increases in potential to be additive. (c) 2010 Elsevier Inc. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据