title: Live cells imaging using a turn-on FRET-based BODIPY probe for biothiols.
abstract: We designed a red-emitting turn-on FRET-based molecular probe 1 for selective detection of cysteine and homocysteine. Probe 1 shows significant fluorescence enhancement after cleavage of the 2, 4-dinitrobenzensulfonyl (DNBS) unit from the fluorophore upon thiols treatment. The precursor of probe 1, BNM153, is a moderate quantum yield FRET dye which contributes a minimum emission leakage from its donor part. We synthesized this assembly by connecting a low quantum yield (less than 1%) BODIPY donor to a high quantum yield BODIPY acceptor via a 1, 3-triazine bridge system. It is noteworthy that the majority of the non-radiative energy loss of donor (BDN) was converted to the acceptor (BDM)'s fluorescence output with minimum leaks of donor emission. The fluorescence sensing mechanism of probe 1 was illustrated by fluorescence spectroscopy, kinetic measurements, HPLC-MS analysis and DFT calculations. Probe 1 is pH-independent at the physiological pH range. Finally, live cells imaging demonstrated the utility of probe 1 as a biosensor for thiols.
authors.affiliation: Department of Chemistry and MedChem Program, Life Sciences Institute, National University of Singapore, 3 Science Drive 3, Singapore 117543, Singapore.
authors.forename: Dongdong
authors.initials: D
authors.lastname: Su
authors.affiliation: Department of Chemistry and MedChem Program, Life Sciences Institute, National University of Singapore, 3 Science Drive 3, Singapore 117543, Singapore.
authors.forename: Chai Lean
authors.initials: CL
authors.lastname: Teoh
authors.affiliation: Department of Chemistry and MedChem Program, Life Sciences Institute, National University of Singapore, 3 Science Drive 3, Singapore 117543, Singapore.
authors.forename: Srikanta
authors.initials: S
authors.lastname: Sahu
authors.affiliation: Department of Chemistry and MedChem Program, Life Sciences Institute, National University of Singapore, 3 Science Drive 3, Singapore 117543, Singapore.
authors.forename: Raj Kumar
authors.initials: RK
authors.lastname: Das
authors.affiliation: Department of Chemistry and MedChem Program, Life Sciences Institute, National University of Singapore, 3 Science Drive 3, Singapore 117543, Singapore; Laboratory of Bioimaging Probe Development, Singapore Bioimaging Consortium (SBIC), 11 Biopolis Way, #02-02 Helios, Agency for Science, Technology and Research (A*STAR), Biopolis, Singapore 138667, Singapore. Electronic address: chmcyt@nus.edu.sg.
authors.forename: Young-Tae
authors.initials: YT
authors.lastname: Chang
mesh_terms: D001896:Boron Compounds / Q000032:analysis* / Q000737:chemistry*; D003545:Cysteine / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006367:HeLa Cells; D006710:Homocysteine / Q000378:metabolism*; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods; D057054:Molecular Imaging / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013438:Sulfhydryl Compounds / Q000378:metabolism*
chemical_list: C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D013438:Sulfhydryl Compounds; D006710:Homocysteine; D003545:Cysteine
keywords: Biothiols; Fluorescence; Fluorescence resonance energy transfer (FRET); Sensor; cysteine