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Site-Specific Fragmentation of Green Fluorescent Protein Induced by Blue Light
Biochemistry ( IF 2.9 ) Pub Date : 2021-07-27 , DOI: 10.1021/acs.biochem.1c00248
Philipp J Heckmeier 1 , Dieter Langosch 1
Affiliation  

Green fluorescent protein (GFP) and related fluorescent proteins have multiple applications in cell biology, and elucidating their functions has been at the focus of biophysical research for about three decades. Fluorescent proteins can be bleached by intense irradiation, and a number of them undergo photoconversion. Rare cases have been reported where distant functional relatives of GFP exhibit UV-light-induced protein fragmentation. Here, we show that irreversible bleaching of two different variants of GFP (sfGFP, EGFP) with visible light is paralleled by successive backbone fragmentation of the protein. Mass spectrometry revealed that the site of fragmentation resides at the fluorophore, between residue positions 65 and 66.

中文翻译:

蓝光诱导绿色荧光蛋白的定点片段化

绿色荧光蛋白 (GFP) 和相关的荧光蛋白在细胞生物学中有多种应用,并且阐明它们的功能一直是生物物理学研究的焦点大约三十年来。荧光蛋白可以通过强辐射漂白,其中一些会发生光转换。据报道,GFP 的远亲功能亲属表现出紫外线诱导的蛋白质断裂的罕见情况。在这里,我们展示了两种不同的 GFP 变体(sfGFP、EGFP)在可见光下的不可逆漂白与蛋白质的连续骨架断裂平行。质谱表明,断裂位点位于荧光团,残基位置 65 和 66 之间。
更新日期:2021-08-17
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