Nienhaus G Ulrich, Nienhaus Karin, Hölzle Angela, Ivanchenko Sergey, Renzi Fabiana, Oswald Franz, Wolff Michael, Schmitt Florian, Röcker Carlheinz, Vallone Beatrice, Weidemann Wolfgang, Heilker Ralf, Nar Herbert, Wiedenmann Jörg
Department of Biophysics, University of Ulm, Ulm, Germany.
Photochem Photobiol. 2006 Mar-Apr;82(2):351-8. doi: 10.1562/2005-05-19-RA-533.
EosFP is a fluorescent protein from the coral Lobophyllia hemprichii that changes its fluorescence emission from green to red upon irradiation with near-UV light. Here we present the spectroscopic properties of wild-type EosFP and a variety of monomeric and dimeric mutants and provide a structural interpretation of its oligomerization and photoconversion, which is based on X-ray structure analysis of the green and red species that we reported recently. Because functional expression of the monomeric EosFP variant is limited to temperatures of 30 degrees C, we have developed a tandem dimer. This construct, in which two EosFP subunits are connected by a flexible 12 amino acid linker, expresses well after fusion with the androgen and endothelin A receptors at 37 degrees C. A variety of applications in cellular imaging, developmental biology and automated high-content screening applications are presented, which demonstrate that EosFP is a powerful tool for in vivo monitoring of cellular processes.
EosFP是一种来自珊瑚赫氏叶状珊瑚（Lobophyllia hemprichii）的荧光蛋白，在用近紫外光照射时，其荧光发射会从绿色变为红色。在此，我们展示了野生型EosFP以及多种单体和二聚体突变体的光谱特性，并基于我们最近报道的绿色和红色物种的X射线结构分析，对其寡聚化和光转换进行了结构解释。由于单体EosFP变体的功能性表达仅限于30摄氏度的温度，我们开发了一种串联二聚体。在这种构建体中，两个EosFP亚基通过一个灵活的12个氨基酸的接头连接，在与雄激素和内皮素A受体在37摄氏度融合后表达良好。本文还介绍了其在细胞成像、发育生物学和自动化高内涵筛选应用中的多种应用，这些应用表明EosFP是用于体内监测细胞过程的强大工具。
