title: Genetically encoded red fluorescent copper(I) sensors for cellular copper(I) imaging.
abstract: Copper ranks among the most important metal ions in living organism, owing to its key catalytic effect in a range of biochemical processes. Dysregulation of in vivo copper(I) metabolism is extremely toxic and would cause serious diseases in human, such as Wilson's and Menkes. Thus, it would be highly valuable to have a proper approach to monitor the dynamics of copper(I) in vivo, as it is directly related to the onset of human copper(I)-related diseases. Under these circumstance, developing fluorescent protein based copper(I) sensors is highly demanded. However, these established sensors are mostly based on green or yellow FPs. Fluorescent copper(I) sensors with a spectra in the red range are more desirable due to lower phototoxicity, less auto-fluorescent noise and better penetration of red light. In the present work, we grafted a special red FP into three different location of a copper(I) binding protein, and generate a series of red fluorescent copper(I) sensors. Despite their limited in vivo sensitivity toward copper(I), these sensors are viable for cellular copper(I) imaging. Furthermore, these red fluorescent copper(I) sensors are a good starting point to develop superior copper(I) biosensors capable of imaging copper(I) fluctuations within a truly biologically relevant concentration, and further effort to realize this endeavor is under way.
authors.affiliation: National Laboratory of Solid State Microstructure, Department of Physics, Nanjing University, Nanjing, Jiangsu 210008, China. Electronic address: jyliang@biophy.nju.edu.cn.
authors.forename: Junyi
authors.initials: J
authors.lastname: Liang
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu 210008, China.
authors.forename: Lele
authors.initials: L
authors.lastname: Guo
authors.affiliation: Sate Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, Jiangsu 210093, China.
authors.forename: Yin
authors.initials: Y
authors.lastname: Ding
authors.affiliation: National Laboratory of Solid State Microstructure, Department of Physics, Nanjing University, Nanjing, Jiangsu 210008, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Xia
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu 210008, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Shen
authors.affiliation: National Laboratory of Solid State Microstructure, Department of Physics, Nanjing University, Nanjing, Jiangsu 210008, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Qin
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu 210008, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Xu
authors.affiliation: National Laboratory of Solid State Microstructure, Department of Physics, Nanjing University, Nanjing, Jiangsu 210008, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Cao
authors.affiliation: National Laboratory of Solid State Microstructure, Department of Physics, Nanjing University, Nanjing, Jiangsu 210008, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
mesh_terms: D015374:Biosensing Techniques; D002477:Cells / Q000378:metabolism*; D003300:Copper / Q000032:analysis*; D005821:Genetic Techniques; D057809:HEK293 Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics*; D057054:Molecular Imaging / Q000379:methods*; D000097573:Red Fluorescent Protein
chemical_list: D003300:Copper; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein
keywords: Biosensor design; Copper(I) sensor; Fluorescent protein; Protein engineering