Richmond T A, Takahashi T T, Shimkhada R, Bernsdorf J
Joint Science Department, Claremont McKenna, Pitzer, and Scripps Colleges, 925 North Mills Avenue, Claremont, California 91711, USA.
Biochem Biophys Res Commun. 2000 Feb 16;268(2):462-5. doi: 10.1006/bbrc.1999.1244.
The ability to assay a variety of metals by noninvasive methods has applications in both biomedical and environmental research. Green fluorescent protein (GFP) is a protein isolated from coelenterates that exhibits spontaneous fluorescence. GFP does not require any exogenous cofactors for fluorescence, and can be easily appended to other proteins at the DNA level, producing a fluorescence-labeled target protein in vivo. Metals in close proximity to chromophores are known to quench fluorescence in a distance-dependent fashion. Potential metal binding sites on the surface of GFP have been identified and mutant proteins have been designed, created, and characterized. These metal-binding mutants of GFP exhibit fluorescence quenching at lower transition metal ion concentrations than those of the wild-type protein. These GFP mutants represent a new class of protein-based metal sensors.
通过非侵入性方法检测多种金属的能力在生物医学和环境研究中均有应用。绿色荧光蛋白（GFP）是一种从腔肠动物中分离出来的能自发荧光的蛋白质。GFP的荧光不需要任何外源辅因子，并且可以在DNA水平上轻松地与其他蛋白质连接，从而在体内产生荧光标记的靶蛋白。已知靠近发色团的金属会以距离依赖的方式淬灭荧光。已鉴定出GFP表面的潜在金属结合位点，并设计、构建和表征了突变蛋白。与野生型蛋白相比，这些GFP的金属结合突变体在较低的过渡金属离子浓度下就表现出荧光淬灭。这些GFP突变体代表了一类新型的基于蛋白质的金属传感器。
