Structural Origins of Altered Spectroscopic Properties upon Ligand Binding in Proteins Containing a Fluorescent Noncanonical Amino Acid.
Structural Origins of Altered Spectroscopic Properties upon Ligand Binding in Proteins Containing a Fluorescent Noncanonical Amino Acid.
复制标题
含有荧光非规范氨基酸的蛋白质中配体结合后光谱特性改变的结构起源。
DOI：
10.1021/acs.biochem.1c00291
复制
发表时间：
2021-08-31
期刊：
影响因子：
2.9
通讯作者：
Mills JH
中科院分区：
文献类型：
--
作者：
Gleason PR;Kolbaba-Kartchner B;Henderson JN;Stahl EP;Simmons CR;Mills JH
荧光非典型氨基酸（fNCAAs）可以作为起点，合理设计的蛋白质为基础的生物活性的荧光传感器。然而，实现这一目标的努力可能受到缺乏蛋白质内fNCAAs原子水平表征的阻碍。在这里，我们描述了5个链霉亲和素突变体，含有fNCAA L-（7-羟基香豆素-4-基）乙基甘氨酸（7-HCAA）在其底物，生物素的结合位点附近的网站的光谱和结构表征。许多突变体显示出响应于生物素结合的改变的荧光光谱，其包括荧光强度的增加和减少以及发射最大值的红移或蓝移。还获得了五个突变体中的三个的结构数据。晶体结构揭示了7-HCAA和功能基团之间的相互作用-由蛋白质或底物贡献-这可能是7-HCAA光谱中观察到的变化的原因。这些数据可用于未来的研究，旨在合理设计的荧光，基于蛋白质的传感器的小分子结合或解离。
Fluorescent non-canonical amino acids (fNCAAs) could serve as starting points for the rational design of protein-based fluorescent sensors of biological activity. However, efforts toward this goal are likely hampered by a lack of atomic-level characterization of fNCAAs within proteins. Here, we describe the spectroscopic and structural characterization of five streptavidin mutants that contain the fNCAA L-(7-hydroxycoumarin-4-yl)ethylglycine (7-HCAA) at sites proximal to the binding site of its substrate, biotin. Many of the mutants exhibited altered fluorescence spectra in response to biotin binding, which included both increases and decreases in fluorescence intensity as well as red or blue shifted emission maxima. Structural data were also obtained for three of the five mutants. The crystal structures shed light on interactions between 7-HCAA and functional groups—contributed either by the protein or substrate—that may be responsible for the observed changes in the 7-HCAA spectra. These data could be used in future studies aimed at the rational design of fluorescent, protein-based sensors of small molecule binding or dissociation.
登录
查看更多内容
DOI：
10.1107/s090744491003982x
发表时间：
2011-04
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Evans PR
通讯作者：
Evans PR
影响因子：
4
作者：
MORIYA, T
通讯作者：
MORIYA, T
影响因子：
15
作者：
Lee HS;Spraggon G;Schultz PG;Wang F
通讯作者：
Wang F
影响因子：
2.9
作者：
Henderson JN;Simmons CR;Fahmi NE;Jeffs JW;Borges CR;Mills JH
通讯作者：
Mills JH
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K