Vogt Alexander, D'Angelo Cecilia, Oswald Franz, Denzel Andrea, Mazel Charles H, Matz Mikhail V, Ivanchenko Sergey, Nienhaus G Ulrich, Wiedenmann Jörg
Institute of General Zoology and Endocrinology, University of Ulm, Ulm, Germany.
PLoS One. 2008;3(11):e3766. doi: 10.1371/journal.pone.0003766. Epub 2008 Nov 19.
A colorful variety of fluorescent proteins (FPs) from marine invertebrates are utilized as genetically encoded markers for live cell imaging. The increased demand for advanced imaging techniques drives a continuous search for FPs with new and improved properties. Many useful FPs have been isolated from species adapted to sun-flooded habitats such as tropical coral reefs. It has yet remained unknown if species expressing green fluorescent protein (GFP)-like proteins also exist in the darkness of the deep sea. Using a submarine-based and -operated fluorescence detection system in the Gulf of Mexico, we discovered ceriantharians emitting bright green fluorescence in depths between 500 and 600 m and identified a GFP, named cerFP505, with bright fluorescence emission peaking at 505 nm. Spectroscopic studies showed that approximately 15% of the protein bulk feature reversible ON/OFF photoswitching that can be induced by alternating irradiation with blue und near-UV light. Despite being derived from an animal adapted to essentially complete darkness and low temperatures, cerFP505 maturation in living mammalian cells at 37 degrees C, its brightness and photostability are comparable to those of EGFP and cmFP512 from shallow water species. Therefore, our findings disclose the deep sea as a potential source of GFP-like molecular marker proteins.
来自海洋无脊椎动物的多种彩色荧光蛋白(FPs)被用作活细胞成像的基因编码标记。对先进成像技术需求的增加推动了对具有新的和改进特性的荧光蛋白的持续探索。许多有用的荧光蛋白已从适应阳光充足栖息地的物种中分离出来,如热带珊瑚礁。目前尚不清楚在深海黑暗环境中是否也存在表达绿色荧光蛋白(GFP)样蛋白的物种。我们在墨西哥湾使用基于潜艇操作的荧光检测系统,发现了在500至600米深度发出亮绿色荧光的海葵,并鉴定出一种名为cerFP505的绿色荧光蛋白,其明亮荧光发射峰值在505纳米。光谱研究表明,大约15%的蛋白具有可逆的开/关光开关特性,可通过交替照射蓝光和近紫外光诱导。尽管cerFP505来源于适应基本完全黑暗和低温环境的动物,但它在37摄氏度的活哺乳动物细胞中能够成熟,其亮度和光稳定性与来自浅水物种的增强型绿色荧光蛋白(EGFP)和cmFP512相当。因此,我们的研究结果揭示了深海是绿色荧光蛋白样分子标记蛋白的潜在来源。