Villa Jordan K, Tran Hong-Anh, Vipani Megha, Gianturco Stephanie, Bhasin Konark, Russell Brent L, Harbron Elizabeth J, Young Douglas D
Department of Chemistry, The College of William & Mary, Williamsburg, VA 231871, USA.
Molecules. 2017 Jul 16;22(7):1194. doi: 10.3390/molecules22071194.
The ability to modulate protein function through minimal perturbations to amino acid structure represents an ideal mechanism to engineer optimized proteins. Due to the novel spectroscopic properties of green fluorescent protein, it has found widespread application as a reporter protein throughout the fields of biology and chemistry. Using site-specific amino acid mutagenesis, we have incorporated various fluorotyrosine residues directly into the fluorophore of the protein, altering the fluorescence and shifting the pKa of the phenolic proton associated with the fluorophore. Relative to wild type GFP, the fluorescence spectrum of the protein is altered with each additional fluorine atom, and the mutant GFPs have the potential to be employed as pH sensors due to the altered electronic properties of the fluorine atoms.
通过对氨基酸结构进行最小程度的扰动来调节蛋白质功能的能力，是构建优化蛋白质的理想机制。由于绿色荧光蛋白具有新颖的光谱特性，它已在生物学和化学领域作为报告蛋白得到广泛应用。利用位点特异性氨基酸诱变技术，我们已将各种氟代酪氨酸残基直接引入蛋白质的荧光团中，改变了荧光并使与荧光团相关的酚性质子的pKa发生了偏移。相对于野生型绿色荧光蛋白，每增加一个氟原子，蛋白质的荧光光谱就会发生改变，并且由于氟原子电子性质的改变，突变型绿色荧光蛋白有潜力用作pH传感器。
