Reaction-based genetically encoded fluorescent hydrogen sulfide sensors.
The detection of hydrogen sulfide (H(2)S), a toxic gas and an important biological signaling molecule, has been a long-time challenge. Here we report genetically encoded fluorescent protein (FP)-based probes that can selectively detect H(2)S. By expanding the genetic codes of E. coli and mammalian cells, FP chromophores were modified with the sulfide-reactive azide functional group. These structurally modified chromophores were selectively reduced by H(2)S, resulting in sensitive fluorescence enhancement detectable by spectroscopic and microscopic techniques. Exploration of a circularly permuted FP led to an improved sensor with faster responses, and the feasibility of using such a genetically encoded probe to monitor H(2)S in living mammalian cells has also been demonstrated.
Department of Chemistry, University of California, 501 Big Springs Road, Riverside, California 92521, USA.
Si
S
Chen
Zhi-jie
ZJ
Chen
Wei
W
Ren
Hui-wang
HW
Ai
D000818:Animals; D001386:Azides / Q000737:chemistry* / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics*; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D006862:Hydrogen Sulfide / Q000032:analysis*; D008958:Models, Molecular; D010084:Oxidation-Reduction
D001386:Azides; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D006862:Hydrogen Sulfide