4.4 Article

Chromophorylation of a Novel Cyanobacteriochrome GAF Domain from Spirulina and Its Response to Copper Ions

Journal

JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
Volume 31, Issue 2, Pages 265-271

Publisher

KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
DOI: 10.4014/jmb.2009.09048

Keywords

Cyanobacteriochrome; phycoerythrobilin; fluorescence quantum yield; biosensor; copper ions; fluorescence quenching

Funding

  1. Natural Science Foundation of Jiangsu Province, China [BK20150884]
  2. National Natural Science Foundation of China [41977354, 41471191]
  3. China Postdoctoral Science Foundation [14ZR1401500]
  4. Jiangsu Planned Projects for Postdoctoral Research Funds [1501020A]
  5. Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)

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CBCRs are phytochrome-related photoreceptor proteins in cyanobacteria covering a wide spectral range with potential applications as biomarkers and biosensors. A novel PEB-binding CBCR GAF domain was identified, exhibiting intense orange fluorescence with high quantum yield, selective and instantaneous quenching by copper ions, and reversibility upon treatment with EDTA, suggesting its potential as a biosensor for rapid copper ion detection.
Cyanobacteriochromes (CBCRs) are phytochrome-related photoreceptor proteins in cyanobacteria and cover a wide spectral range from ultraviolet to far-red. A single GAF domain that they contain can bind bilin(s) autocatalytically via heterologous recombination and then fluoresce, with potential applications as biomarkers and biosensors. Here, we report that a novel red/green CBCR GAF domain, SPI1085g2 from Spirulina subsalsa, covalently binds both phycocyanobilin (PCB) and phycoerythrobilin (PEB). The PCB-binding GAF domain exhibited canonical red/green photoconversion with weak fluorescence emission. However, the PEB-binding GAF domain, SPI1085g2-PEB, exhibited an intense orange fluorescence(lambda(abs.max)= 520 nm, lambda(fluor. max)= 555 nm), with a fluorescence quantum yield close to 1.0. The fluorescence of SPI1085g2-PEB was selectively and instantaneously quenched by copper ions in a concentration-dependent manner and exhibited reversibility upon treatment with the metal chelator EDTA. This study identified a novel PEB-binding cyanobacteriochrome-based fluorescent protein with the highest quantum yield reported to date and suggests its potential as a biosensor for the rapid detection of copper ions.

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