A versatile amino acid analogue of the solvatochromic fluorophore 4-N,N-dimethylamino-1,8-naphthalimide: a powerful tool for the study of dynamic protein interactions.
A versatile amino acid analogue of the solvatochromic fluorophore 4-N,N-dimethylamino-1,8-naphthalimide: a powerful tool for the study of dynamic protein interactions.
复制标题
DOI：
10.1021/ja804754y
复制
发表时间：
2008-10-15
影响因子：
15
通讯作者：
Imperiali B
中科院分区：
文献类型：
--
作者：
Loving G;Imperiali B
我们开发了一种新的非天然氨基酸的基础上的溶剂化显色荧光团4-N，N-二甲氨基-1，8-萘酰亚胺（4-DMN）的蛋白质-蛋白质相互作用的研究中的应用。该发色团的荧光量子产率对局部溶剂环境的变化高度敏感，证明了荧光团的二甲基氨基邻苯二甲酰亚胺家族的“开关样”发射特性。特别是，这种新的物种具有许多显着的优势，相关的荧光团，包括更大的化学稳定性，在广泛的条件下，更长的激发波长（408 nm），并改善合成的可及性。该氨基酸已被制备为Fmoc保护的结构单元，并且可以通过标准固相肽合成（SPPS）容易地掺入肽中。一系列的比较研究表明，4-DMN氨基酸相对于以前报道的4-N，N-二甲基氨基邻苯二甲酰亚胺丙氨酸（4-达帕）和6-N，N-二甲基氨基-2，3-萘二甲酰亚胺丙氨酸（6-DMNA）氨基酸的有利性质。其他市售溶剂化显色荧光团也包括在这些研究中。这种新的探针作为研究蛋白质-蛋白质相互作用的工具的潜力是通过将其引入到由钙激活的钙调蛋白识别的肽中来证明的。这两种组分之间的结合相互作用产生大于900倍的荧光发射增加。
We have developed a new unnatural amino acid based on the solvatochromic fluorophore 4-N,N-dimethylamino-1,8-naphthalimide (4-DMN) for application in the study of protein-protein interactions. The fluorescence quantum yield of this chromophore is highly sensitive to changes in the local solvent environment, demonstrating “switch-like” emission properties characteristic of the dimethylaminophthalimide family of fluorophores. In particular, this new species possesses a number of significant advantages over related fluorophores including greater chemical stability under a wide range of conditions, a longer wavelength of excitation (408 nm), and improved synthetic accessibility. This amino acid has been prepared as an Fmoc-protected building block and may readily be incorporated into peptides via standard solid-phase peptide synthesis (SPPS). A series of comparative studies are presented to demonstrate the advantageous properties of the 4-DMN amino acid relative to that of the previously reported 4-N,N-dimethylaminophthalimidoalanine (4-DAPA) and 6-N,N-dimethylamino-2,3-naphthalimidoalanine (6-DMNA) amino acids. Other commercially available solvatochromic fluorophores are also include in these studies. The potential of this new probe as a tool for the study of protein-protein interactions is demonstrated by introducing it into a peptide that is recognized by calcium-activated calmodulin. The binding interaction between these two components yields an increase in fluorescence emission greater than 900-fold.
登录
查看更多内容
影响因子：
56.9
作者：
IKURA, M;CLORE, GM;BAX, A
通讯作者：
BAX, A
影响因子：
--
作者：
Hachmann, J;Lebl, M
通讯作者：
Lebl, M
影响因子：
1.8
作者：
Dufau, I;Mazarguil, H
通讯作者：
Mazarguil, H
DOI：
10.1039/p29900000837
发表时间：
1990-05-01
期刊：
JOURNAL OF THE CHEMICAL SOCIETY-PERKIN TRANSACTIONS 2
影响因子：
--
作者：
ALEXIOU, MS;TYCHOPOULOS, V;BRITTAIN, PI
通讯作者：
BRITTAIN, PI
影响因子：
7.3
作者：
Berque-Bestel, I;Soulier, JL;Sicsic, S
通讯作者：
Sicsic, S