Photoconvertible fluorescent protein EosFP:: Biophysical properties and cell biology applications

被引:98
作者
Nienhaus, GU [1 ]
Nienhaus, K
Hölzle, A
Ivanchenko, S
Renzi, F
Oswald, F
Wolff, M
Schmitt, F
Röcker, C
Vallone, B
Weidemann, W
Heilker, R
Nar, H
Wiedenmann, J
机构
[1] Univ Ulm, Dept Biophys, Ulm, Germany
[2] Univ Illinois, Dept Phys, Urbana, IL 61801 USA
[3] Univ Roma La Sapienza, Dept Biochem Sci, Rome, Italy
[4] Univ Ulm, Dept Internal Med 1, D-7900 Ulm, Germany
[5] Univ Ulm, Dept Gen Zool & Endocrinol, Ulm, Germany
[6] Boehringer Ingelheim KG, Dept Lead Discovery, Biberach, Germany
关键词
D O I
10.1562/2005-05-19-RA-533
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:351 / 358
页数:8
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