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首页> 外文期刊>Acta crystallographica. Section D, Structural biology. >Crystal structure of the fluorescent protein from Dendronephthya sp in both green and photoconverted red forms
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Crystal structure of the fluorescent protein from Dendronephthya sp in both green and photoconverted red forms

机译：荧光蛋白的晶体结构在绿色和Dendronephthya spphotoconverted红色形式

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摘要

The fluorescent protein from Dendronephthya sp. (DendFP) is a member of the Kaede-like group of photoconvertible fluorescent proteins with a His62-Tyr63-Gly64 chromophore-forming sequence. Upon irradiation with UV and blue light, the fluorescence of DendFP irreversibly changes from green (506 nm) to red (578 nm). The photoconversion is accompanied by cleavage of the peptide backbone at the C-alpha-N bond of His62 and the formation of a terminal carboxamide group at the preceding Leu61. The resulting double C-alpha=C-beta bond in His62 extends the conjugation of the chromophore pi system to include imidazole, providing the red fluorescence. Here, the three-dimensional structures of native green and photoconverted red forms of DendFP determined at 1.81 and 2.14 angstrom resolution, respectively, are reported. This is the first structure of photoconverted red DendFP to be reported to date. The structure-based mutagenesis of DendFP revealed an important role of positions 142 and 193: replacement of the original Ser142 and His193 caused a moderate red shift in the fluorescence and a considerable increase in the photoconversion rate. It was demonstrated that hydrogen bonding of the chromophore to the Gln116 and Ser105 cluster is crucial for variation of the photoconversion rate. The single replacement Gln116Asn disrupts the hydrogen bonding of Gln116 to the chromophore, resulting in a 30-fold decrease in the photoconversion rate, which was partially restored by a further Ser105Asn replacement.
机译：Dendronephthya sp的荧光蛋白。(DendFP)是Kaede-like集团的一员photoconvertible荧光蛋白的His62-Tyr63-Gly64 chromophore-forming序列。在辐照紫外和蓝光荧光DendFP不可逆转的变化绿色(506海里)红色(578海里)。photoconversion伴随着的乳沟肽His62 C-alpha-N债券的支柱和一个终端的形成酰胺组在前面的Leu61。C-alpha = C-beta His62扩展了债券发色团的共轭π体系包括咪唑,提供红荧光。本机结构绿色和photoconverted红色形式的DendFP决定在1.81和2.14埃分辨率分别被报道。这是第一个photoconverted红色的结构DendFP报告。诱变DendFP透露一个小142年和193年重要的职位:替换原来的Ser142 His193造成一个温和的荧光红移和一个相当大的增加photoconversion率。Gln116氢键的发色团和Ser105集群的变化是至关重要的photoconversion率。Gln116Asn扰乱Gln116的氢键发色团,导致30倍减少photoconversion率,这是进一步Ser105Asn部分恢复更换。

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