Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520, USA.
FEBS J. 2010 Apr;277(8):1967-78. doi: 10.1111/j.1742-4658.2010.07618.x. Epub 2010 Mar 18.
Fluorescent proteins have become essential tools in molecular and biological applications. Here, we present a novel fluorescent protein isolated from warm water coral, Cyphastrea microphthalma. The protein, which we named vivid Verde fluorescent protein (VFP), matures readily at 37 degrees C and emits bright green light. Further characterizations revealed that VFP has a tendency to form dimers. By creating a homology model of VFP, based on the structure of the red fluorescent protein, DsRed, we were able to make mutations that alter the protein's oligomerization state. We present two proteins, mVFP and mVFP1, that are both exclusively monomeric, and one protein, dVFP, which is dimeric. We characterized the spectroscopic properties of VFP and its variants in comparison with enhanced green fluorescent protein (EGFP), a widely used variant of GFP. All the VFP variants are at least twice as bright as EGFP. Finally, we demonstrated the effectiveness of the VFP variants in both in vitro and in vivo detection applications.
荧光蛋白已成为分子和生物应用中不可或缺的工具。在这里，我们从暖水珊瑚 Cyphastrea microphthalma 中分离出一种新型荧光蛋白。该蛋白我们命名为 vivid Verde 荧光蛋白 (VFP)，在 37°C 下即可成熟，并发出明亮的绿光。进一步的特性分析表明，VFP 有形成二聚体的趋势。通过基于红色荧光蛋白 DsRed 的结构创建 VFP 的同源模型，我们能够进行突变，从而改变蛋白质的寡聚状态。我们提出了两种蛋白质，mVFP 和 mVFP1，它们都是纯单体，而另一种蛋白质 dVFP 是二聚体。我们比较了 VFP 及其变体与广泛使用的 GFP 变体增强型绿色荧光蛋白 (EGFP) 的光谱特性。所有 VFP 变体的亮度至少是 EGFP 的两倍。最后，我们证明了 VFP 变体在体外和体内检测应用中的有效性。
