A New-Generation Fluorescent-Based Metal Sensor - iLOV Protein

Title
A New-Generation Fluorescent-Based Metal Sensor - iLOV Protein
Author(s)
이종순Yuvaraj Ravikumar[Yuvaraj Ravikumar]Saravanan Prabhu Nadarajan[Saravanan Prabhu Nadarajan]Jin-Kyu Rhee[Jin-Kyu Rhee]Hyungdon Yun[Hyungdon Yun]
Keywords
BIOSENSOR; REPORTER; VARIANTS; IONS; GFP
Issue Date
201504
Publisher
KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
Citation
JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY, v.25, no.4, pp.503 - 510
Abstract
The iLOV protein belongs to a family of blue-light photoreceptor proteins containing a light-oxygen-voltage sensing domain with a noncovalently bound flavin mononucleotide (FMN) as its chromophore. Owing to advantages such as its small size, oxygen-independent nature, and pH stability, iLOV is an ideal candidate over other reporter fluorescent proteins such as GFP and DsRed. Here, for the first time, we describe the feasibility of applying LOV domain-based fluorescent iLOV as a metal sensor by measuring the fluorescence quenching of a protein with respect to the concentration of metal ions. In the present study, we demonstrated the inherent copper sensing property of the iLOV protein and identified the possible amino acids responsible for metal binding. The fluorescence quenching upon exposure to Cu2+ was highly sensitive and exhibited reversibility upon the addition of the metal chelator EDTA. The copper binding constant was found to be 4.72 +/- 0.84 mu M. In addition, Cu2+-bound iLOV showed high fluorescence quenching at near physiological pH. Further computational analysis yielded a better insight into understanding the possible amino acids responsible for Cu2+ binding with the iLOV protein.
URI
http://hdl.handle.net/YU.REPOSITORY/32707http://dx.doi.org/10.4014/jmb.1409.09035
ISSN
1017-7825
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이과대학 > 화학생화학부 > Articles
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