Synthesis of fluorescent dipeptidomimetics and their ribosomal incorporation into green fluorescent protein.
Synthesis of fluorescent dipeptidomimetics and their ribosomal incorporation into green fluorescent protein.
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DOI：
10.1016/j.bmcl.2015.08.073
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发表时间：
2015-11-01
影响因子：
2.7
通讯作者：
Hecht SM
中科院分区：
文献类型：
--
作者：
Roy Chowdhury S;Maini R;Dedkova LM;Hecht SM
描述了通过体外蛋白质翻译的基于新的恶唑和噻唑的二肽模拟物的合成和掺入到绿色荧光蛋白（GFP）的66位。这些化合物可以被认为是GFP发色团类似物，并且是强荧光的。保护的甘氨酸经双酰化得到α-氨基-β-酮酯中间体。将中间体脱水环化得到1，3-恶唑，用Lawesson试剂处理得到1，3-噻唑。当将这些荧光团引入GFP的66位取代Tyr 66时，所得GFP类似物表现出比野生型GFP大几倍的荧光发射;发射也向更短的波长移动。可以注意到，与天然和修饰的荧光蛋白中形成的典型荧光团相比，恶唑和噻唑荧光团是完全稳定的，并且不需要通过翻译后修饰来活化以显示荧光。
The synthesis and incorporation into position 66 of green fluorescent protein (GFP) by in vitro protein translation of novel oxazole and thiazole based dipeptidomimetics are described. The compounds may be regarded as GFP chromophore analogues, and are strongly fluorescent. An α-amido-β-ketoester intermediate was obtained via bisacylation of a protected glycine. The intermediate underwent dehydrative cyclization to afford the 1,3-oxazole and was treated with Lawesson’s reagent to furnish the 1,3-thiazole. When these fluorophores were introduced into position 66 of GFP in place of Tyr66, the resulting GFP analogues exhibited fluorescence emission several-fold greater than wild-type GFP; the emission was also shifted to shorter wavelength. It may be noted that compared to the typical fluorophores formed in the natural and modified fluorescent proteins, the oxazole and thiazole fluorophores are completely stable and do not require activation by posttranslational modification to exhibit fluorescence.