In vivo targeting of organic calcium sensors via genetically selected peptides
In vivo targeting of organic calcium sensors via genetically selected peptides
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DOI：
10.1016/j.chembiol.2004.03.004
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发表时间：
2004-03-01
影响因子：
--
通讯作者：
Nolan, GP
中科院分区：
文献类型：
--
作者：
Marks, KM;Rosinov, M;Nolan, GP
筛选形成稳定折叠结构的受约束肽的文库中以高亲和力结合荧光染料德克萨斯红的适体。两个选择的克隆具有25和80 pM的噬菌体的德克萨斯红的结合常数，并且结合对德克萨斯红的荧光具有最小的影响。这些肽与德克萨斯红的不同但重叠的区域相互作用。一种肽与X-rhod钙传感器结合，所述X-rhod钙传感器与德克萨斯红共享相同的核心荧光团。这些染料在与肽结合时保持钙敏感性。该肽用于在体内标记与X-rhod-5 F的融合蛋白，并且X-rhod Sensed局部改变钙。因此，最小的，受约束的肽可以在生物学背景下功能性地结合到环境敏感的染料或其他有机试剂，提示用于体内成像和分析的工具。
A library of constrained peptides that form stable folded structures was screened for aptamers that bind with high affinity to the fluorescent dye Texas red. Two selected clones had binding constants to Texas red of 25 and 80 pM as phage and binding had minimal effects on the fluorescence of Texas red. The peptides interact with distinct but overlapping regions of Texas red. One peptide bound to X-rhod calcium sensors which share the same core fluorophore as Texas red These dyes retained calcium sensitivity when bound to the peptide. This peptide was used to label a fusion protein with X-rhod-5F in vivo, and X-rhod Sensed changes in calcium locally. Thus, minimal, constrained peptides can functionally bind to environmentally sensitive dyes or other organic agents in biological contexts, suggesting tools for in vivo imaging and analysis.