title: Labeling thiols on proteins, living cells, and tissues with enhanced emission induced by FRET.
abstract: Using N-(2-Aminoethyl)maleimide-cysteine(StBu) (Mal-Cys) as a medium, protein thiols were converted into N-terminal cysteines. After a biocompatible condensation reaction between the N-terminal cysteine and fluorescent probe 2-cyanobenzothiazole-Gly-Gly-Gly-fluorescein isothiocyanate (CBT-GGG-FITC), a new fluorogenic structure Luciferin-GGG-FITC was obtained. The latter exhibits near one order of magnitude (7 folds) enhanced fluorescence emission compared to the precursor moiety due to fluorescence resonance energy transfer (FRET) effect between the newly formed luciferin structure and the FITC motif. Theoretical investigations revealed the underlying mechanism that satisfactorily explained the experimental results. With this method, enhanced fluorescence imaging of thiols on proteins, outer membranes of living cells, translocation of membrane proteins, and endothelial cell layers of small arteries was successfully achieved.
authors.affiliation: CAS Key Laboratory of Soft Matter Chemistry, Department of Chemistry, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230026, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Yuan
authors.affiliation: Department of Chemical Physics, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230026, China.
authors.forename: Xijun
authors.initials: X
authors.lastname: Wang
authors.affiliation: CAS Key Laboratory of Soft Matter Chemistry, Department of Chemistry, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230026, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Mei
authors.affiliation: CAS Key Laboratory of Soft Matter Chemistry, Department of Chemistry, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230026, China.
authors.forename: Dongxin
authors.initials: D
authors.lastname: Zhang
authors.affiliation: CAS Key Laboratory of Soft Matter Chemistry, Department of Chemistry, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230026, China.
authors.forename: Anming
authors.initials: A
authors.lastname: Tang
authors.affiliation: CAS Key Laboratory of Soft Matter Chemistry, Department of Chemistry, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230026, China.
authors.forename: Linna
authors.initials: L
authors.lastname: An
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082, China.
authors.forename: Xiaoxiao
authors.initials: X
authors.lastname: He
authors.affiliation: 1] Department of Chemical Physics, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230026, China [2] Guizhou Provincial Key Laboratory of Computational Nano-Material Science, Guizhou Normal College, Guiyang 550018, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Jiang
authors.affiliation: 1] CAS Key Laboratory of Soft Matter Chemistry, Department of Chemistry, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230026, China [2] State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082, China.
authors.forename: Gaolin
authors.initials: G
authors.lastname: Liang
mesh_terms: D000818:Animals; D001921:Brain; D002462:Cell Membrane / Q000737:chemistry; D003545:Cysteine / Q000031:analogs & derivatives / Q000737:chemistry; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes; D008301:Maleimides / Q000737:chemistry; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*; D013438:Sulfhydryl Compounds / Q000737:chemistry*
chemical_list: D005456:Fluorescent Dyes; D008301:Maleimides; D008565:Membrane Proteins; D013438:Sulfhydryl Compounds; C062020:S-(bismaleimidoethyl ether)cysteine; D012710:Serum Albumin, Bovine; D016650:Fluorescein-5-isothiocyanate; D003545:Cysteine