Synthesis and properties of the red chromophore of the green-to-red photoconvertible fluorescent protein Kaede and its analogs.
Synthesis and properties of the red chromophore of the green-to-red photoconvertible fluorescent protein Kaede and its analogs.
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DOI：
10.1016/j.bioorg.2007.12.003
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发表时间：
2008-04
影响因子：
5.1
通讯作者：
I. Yampolsky;A. Kislukhin;Tynchtyk Amatov;D. Shcherbo;V. Potapov;S. Lukyanov;K. Lukyanov
中科院分区：
文献类型：
--
作者：
I. Yampolsky;A. Kislukhin;Tynchtyk Amatov;D. Shcherbo;V. Potapov;S. Lukyanov;K. Lukyanov
绿色荧光蛋白（GFP）及其同源蛋白具有独特的基于自身氨基酸残基的自催化修饰的发色团形成途径。绿色到红色光转换荧光蛋白Kaede携带His-Tyr-Gly发色团形成三联体。在这里，我们描述了Kaede红发色团（2-[（1 E）-2-（5-咪唑基）乙烯基]-4-（对羟基苯亚甲基）-5-咪唑酮）及其类似物的合成，这些类似物可以由天然氨基酸残基潜在地形成。获得对应于以下三肽的发色团：His-Tyr-Gly、Trp-Tyr-Gly、Phe-Trp-Gly、Tyr-Trp-Gly、Asn-Tyr-Gly、Phe-Tyr-Gly和Tyr-Tyr-Gly。在碱性条件下，它们以相对高的量子产率（对于Trp衍生的化合物高达0.017）发出红色荧光。在碱性DMSO中，Trp-Tyr-Gly的吸收峰在595 nm处发生红移。令人惊讶的是，在碱性DMF中，非芳族Asn衍生的发色团Asn-Tyr-Gly在642 nm处表现出最大红移发射。因此，Asn残基可能是一个很有前途的取代基，它可能会使GFP样蛋白的翻译后化学多样化。
Green fluorescent protein (GFP) and homologous proteins possess a unique pathway of chromophore formation based on autocatalytic modification of their own amino acid residues. Green-to-red photoconvertible fluorescent protein Kaede carries His–Tyr–Gly chromophore-forming triad. Here, we describe synthesis of Kaede red chromophore (2-[(1E)-2-(5-imidazolyl)ethenyl]-4-(p-hydroxybenzylidene)-5-imidazolone) and its analogs that can be potentially formed by natural amino acid residues. Chromophores corresponding to the following tripeptides were obtained: His–Tyr–Gly, Trp–Tyr–Gly, Phe–Trp–Gly, Tyr–Trp–Gly, Asn–Tyr–Gly, Phe–Tyr–Gly, and Tyr–Tyr–Gly. In basic conditions they fluoresced red with relatively high quantum yield (up to 0.017 for Trp-derived compounds). The most red-shifted absorption peak at 595nm was found for the chromophore Trp–Tyr–Gly in basic DMSO. Surprisingly, in basic DMF non-aromatic Asn-derived chromophore Asn–Tyr–Gly demonstrated the most red-shifted emission maximum at 642nm. Thus, Asn residue may be a promising substituent, which can potentially diversify posttranslational chemistry in GFP-like proteins.