Non–fluorescent mutant of green fluorescent protein sheds light on the mechanism of chromophore formation

Bartkiewicz Ma?gorzata; Kazazi? Sa?a; Krasowska Joanna; Clark Patricia L.; Wielgus‐Kutrowska Beata; Bzowska Agnieszka

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Non–fluorescent mutant of green fluorescent protein sheds light on the mechanism of chromophore formation

机译：绿色荧光蛋白的非荧光突变体揭示了发色团形成的机制

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The mechanism of green fluorescent protein ( GFP ) chromophore formation is still not clearly defined. Two mechanisms have been proposed: cyclisation–dehydration–oxidation (Mechanism A) and cyclisation–oxidation–dehydration (Mechanism B). To distinguish between these mechanisms, we generated a non–fluorescent mutant of GFP , S65T/G67A‐ GFP . This mutant folds to a stable, native‐like structure but lacks fluorescence due to interruption of the chromophore maturation process. Mass spectrometric analysis of peptides derived from this mutant reveal that chromophore formation follows only mechanism A, but that the final oxidation reaction is suppressed. This result is unexpected within the pool of examined GFP mutants, since for the wild‐type GFP , there is strong support for mechanism B.

机译：仍未明确定义绿色荧光蛋白（GFP）发色团形成的机制。已经提出了两种机制：环化 - 脱水 - 氧化（机制A）和环化 - 氧化 - 脱水（机制B）。为了区分这些机制，我们产生了GFP，S65T / G67A-GFP的非荧光突变体。该突变体折叠成稳定的天然样结构，但由于发色团成熟过程的中断，缺乏荧光。源自该突变体衍生的肽的质谱分析揭示了发色团形成遵循机制A，而是抑制了最终氧化反应。该结果在检查的GFP突变体池中出现意外，因为对于野生型GFP，机制B的强大支持。

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来源
《FEBS letters.》 |2018年第9期|共8页
作者
Bartkiewicz Ma?gorzata; Kazazi? Sa?a; Krasowska Joanna; Clark Patricia L.; Wielgus‐Kutrowska Beata; Bzowska Agnieszka;
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作者单位

Division of BiophysicsUniversity of WarsawPoland;

Institut Ru?er Bo?kovi?Zagreb Croatia;

Division of BiophysicsUniversity of WarsawPoland;

Department of Chemistry &

BiochemistryUniversity of Notre DameIN USA;

Division of BiophysicsUniversity of WarsawPoland;

Division of BiophysicsUniversity of WarsawPoland;

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原文格式 PDF
正文语种 eng
中图分类生物化学;
关键词
chromophore maturation; green fluorescent protein; ms/ms spectrometry; S65T mutant; S65T/G67A mutant;

机译：发色团成熟;绿色荧光蛋白;MS / MS光谱;S65T突变体;S65T / G67A突变体;

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专利

1. Non–fluorescent mutant of green fluorescent protein sheds light on the mechanism of chromophore formation [J] . Magorzata Bartkiewicz, Saa Kazazi, Joanna Krasowska, FEBS Letters . 2018,第9期

机译：绿色荧光蛋白的非荧光突变体阐明了生色团的形成机理
2. Shedding light on the dark and weakly fluorescent states of green fluorescent proteins [J] . WOLFGANG WEBER, J. ANDRE MCCAMMON Proceedings of the National Academy of Sciences of the United States of America . 1999,第11期

机译：在绿色荧光蛋白的黑暗和弱荧光状态下发光
3. Ab initio molecular dynamics of the green fluorescent protein (GFP)Chromophore:an insight into the photoinduced dynamics of green fluorescent proteins [J] . Valetina Tozzini, Riccardo Nifosi The journal of physical chemistry, B. Condensed matter, materials, surfaces, interfaces & biophysical . 2001,第24期

机译：从头算分子动力学的绿色荧光蛋白（GFP）发色团：绿色荧光蛋白的光诱导动力学的见解
4. Multi-pixel spectral imaging of green fluorescent protein (GFP) in COS-7 cells: folding kinetics and chromophore formation [C] . Lior Greenbaum, Chana Rothmann, Judith Hanania, Conference on Laser Microscopy, Jul 7-8, 2000, Amsterdam, Netherlands . 2000

机译：COS-7细胞中绿色荧光蛋白（GFP）的多像素光谱成像：折叠动力学和生色团形成
5. Conformational states and transitions in green fluorescent protein chromophore studied by single molecule SERS. [D] . Khurshid, Md Shafayet. 2011

机译：通过单分子SERS研究的绿色荧光蛋白发色团的构象状态和跃迁。
6. Shedding light on the dark and weakly fluorescent states of green fluorescent proteins [O] . Wolfgang Weber, Volkhard Helms, J. Andrew McCammon, 1999

机译：在绿色荧光蛋白的黑暗和弱荧光状态下发光
7. Non–fluorescent mutant of green fluorescent protein sheds light on the mechanism of chromophore formation [O] . Małgorzata Bartkiewicz, Saša Kazazić, Joanna Krasowska, 2018

机译：绿色荧光蛋白的非荧光突变体揭示了发色团形成的机制
8. Mechanisms of Formation and Structure of Chromophores of Fluorescent Proteins from Anthoza Species [R] . Lukyanov, S. A. 2005

机译：anthoza物种荧光蛋白发色团的形成和结构机制

Non–fluorescent mutant of green fluorescent protein sheds light on the mechanism of chromophore formation

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