Disrupting Fluorescence by Mutagenesis in a Green Fluorescent Fatty Acid Binding Protein from a Marine Eel
Disrupting Fluorescence by Mutagenesis in a Green Fluorescent Fatty Acid Binding Protein from a Marine Eel
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DOI：
10.1007/s10930-020-09883-3
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发表时间：
2020-02
期刊：
影响因子：
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通讯作者：
Sara R Krivoshik;Andrew M. Guarnaccia;Daniel B. Fried;D. F. Gruber;Jean P. Gaffney
中科院分区：
文献类型：
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作者：
Sara R Krivoshik;Andrew M. Guarnaccia;Daniel B. Fried;D. F. Gruber;Jean P. Gaffney
人们发现生物荧光是海洋中越来越普遍的现象。隐居的加勒比海鳗鱼 Kaupichthys hyoproroides 在其原生海洋环境中显示出明亮的绿色荧光。我们之前已经证明荧光归因于荧光脂肪酸结合蛋白 Chlopsid FP，它是荧光脂肪酸结合蛋白大家族的一部分，包括同源的 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 Chlopsid FP 荧光猝灭的缺失。此外，我们报告了通过荧光滴定测量的胆红素与 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的荧光潜在机制提供了重要见解。
Biofluorescence has been found to be an increasingly widespread phenomenon in the ocean. The reclusive Caribbean chlopsid eel,Kaupichthys hyoproroidesdisplays 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.