Hoi Hiofan, Howe Elizabeth S, Ding Yidan, Zhang Wei, Baird Michelle A, Sell Brittney R, Allen John R, Davidson Michael W, Campbell Robert E
Department of Chemistry, University of Alberta, Edmonton, AB T6G 2G2, Canada.
Chem Biol. 2013 Oct 24;20(10):1296-304. doi: 10.1016/j.chembiol.2013.08.008. Epub 2013 Oct 3.
Protein engineering has created a palette of monomeric fluorescent proteins (FPs), but there remains an ~30 nm spectral gap between the most red-shifted useful Aequorea victoria green FP (GFP) variants and the most blue-shifted useful Discosoma sp. red FP (RFP) variants. To fill this gap, we have engineered a monomeric version of the yellow FP (YFP) from Zoanthus sp. coral. Our preferred variant, designated as mPapaya1, displays excellent fluorescent brightness, good photostability, and retains its monomeric character both in vitro and in living cells in the context of protein chimeras. We demonstrate that mPapaya1 can serve as a good Förster resonance energy transfer (FRET) acceptor when paired with an mTFP1 donor. mPapaya1 is a valuable addition to the palette of FP variants that are useful for multicolor imaging and FRET-based biosensing.
蛋白质工程已经创造了一系列单体荧光蛋白（FPs），但是在最红移的有用的维多利亚多管水母绿色荧光蛋白（GFP）变体和最蓝移的有用的盘状珊瑚红色荧光蛋白（RFP）变体之间仍然存在约30纳米的光谱间隙。为了填补这一间隙，我们对来自纽扣珊瑚的黄色荧光蛋白（YFP）进行了单体化改造。我们优选的变体命名为mPapaya1，它具有出色的荧光亮度、良好的光稳定性，并且在蛋白质嵌合体的背景下，在体外和活细胞中均保持其单体特性。我们证明，当与mTFP1供体配对时，mPapaya1可以作为良好的荧光共振能量转移（FRET）受体。mPapaya1是对用于多色成像和基于FRET的生物传感的荧光蛋白变体库的一个有价值的补充。
