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Disrupting Fluorescence by Mutagenesis in a Green Fluorescent Fatty Acid Binding Protein from a Marine Eel.
The Protein Journal ( IF 3 ) Pub Date : 2020-02-24 , DOI: 10.1007/s10930-020-09883-3
Sara Rose Krivoshik 1, 2 , Andrew M Guarnaccia 1, 2 , Daniel B Fried 3 , David F Gruber 1, 2 , Jean P Gaffney 1, 2
Affiliation  

Biofluorescence has been found to be an increasingly widespread phenomenon in the ocean. The reclusive Caribbean chlopsid eel, Kaupichthys hyoproroides displays bright green fluorescence in its native marine environment. We have previously shown the fluorescence to be attributed to a fluorescent fatty acid-binding protein, Chlopsid FP, part of a larger family of fluorescent fatty acid-binding proteins, including the homologous UnaG. All require the addition of exogenous bilirubin for fluorescence. Here, we report the generation of a series of point mutants, and deletions that result in the quenching of fluorescence in Chlopsid FP. In addition, we report the binding constants of bilirubin to Chlopsid FP and mutants, measured by fluorescence titration. This study provides key insights into the potential mechanism of fluorescence in this class of fluorescent fatty acid-binding proteins.

中文翻译:

通过诱变破坏来自海鳗的绿色荧光脂肪酸结合蛋白中的荧光。

已经发现生物荧光是海洋中越来越普遍的现象。隐逸的加勒比鱼,Kaupichthys hyoproroides在其原生海洋环境中显示亮绿色的荧光。我们以前已经显示出荧光归因于荧光脂肪酸结合蛋白Chlopsid FP,属于更大的荧光脂肪酸结合蛋白家族,包括同源UnaG。所有这些都需要添加外源性胆红素来产生荧光。在这里,我们报告了一系列点突变体的产生,以及导致Champsid FP中荧光猝灭的缺失。此外,我们报告了通过荧光滴定法测定的胆红素与Chlopsid FP和突变体的结合常数。这项研究为此类荧光脂肪酸结合蛋白中的荧光潜在机理提供了重要见解。
更新日期:2020-02-24
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