A New-Generation Fluorescent-Based Metal Sensor – iLOV Protein
- 주제(키워드) Copper , Fluorescence quenching , iLOV protein , Metal sensor
- 등재 SCIE, KCI등재, SCOPUS
- 발행기관 Korean Society for Microbiolog and Biotechnology
- 발행년도 2015
- 총서유형 Journal
- URI http://www.dcollection.net/handler/ewha/000000115571
- 본문언어 영어
- Published As http://dx.doi.org/10.4014/jmb.1409.09035
- 저작권 이화여자대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
The iLOV protein belongs to a family of blue-light photoreceptor proteins containing a lightoxygen- 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 μM. 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. © 2015 by The Korean Society for Microbiology and Biotechnology.
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