title: A fluorogenic probe for imaging protein S-nitrosylation in live cells.
abstract: S-nitrosylation is a posttranslational modification of protein cysteine residues leading to the formation of S-nitrosothiols and its detection is crucial to understanding of redox regulation and NO-based signaling. Prototypical detection methods for S-nitrosylation are always carried out ex situ. However, the reversible nature and the tendency of transnitrosylation highlight the necessity of its probing in intact live biological contexts. Herein we provide a fluorogenic chemical probe for the detection of S-nitrosylation in live endothelial cells. The probe is weakly emissive alone and becomes highly fluorescent only after undergoing a reaction with S-nitrosothiols in live cellular environments. This probe features high degrees of specificity and desirable sensitivity. Furthermore, it has been successfully applied to image the dynamic change of protein S-nitrosylation in live endothelial cells. The applicability of the probe in complex biological systems has been additionally verified by imaging a known target of S-nitrosylation, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), in live cells. Due to the versatility exemplified, this probe holds great promise for exploring the role of protein S-nitrosylation in the pathophysiological process of a variety of vascular diseases.
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors.forename: Shiyi
authors.initials: S
authors.lastname: Shao
authors.affiliation: Department of Chemistry, Zhejiang Sci-Tech University, Hangzhou 310018, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Chen
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Cheng
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors.forename: Chengkun
authors.initials: C
authors.lastname: Wang
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors.forename: Yanli
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors.forename: Lingxiao
authors.initials: L
authors.lastname: Shao
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
authors.forename: Yongzhou
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Department of Chemistry, Zhejiang Sci-Tech University, Hangzhou 310018, China.
authors.forename: Yifeng
authors.initials: Y
authors.lastname: Han
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: changhuahan@zju.edu.cn.
authors.forename: Feng
authors.initials: F
authors.lastname: Han
authors.affiliation: College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China. Electronic address: lixin81@zju.edu.cn.
authors.forename: Xin
authors.initials: X
authors.lastname: Li
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D003545:Cysteine / Q000737:chemistry; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry*; D006801:Humans; D009569:Nitric Oxide / Q000378:metabolism*; D010084:Oxidation-Reduction; D011499:Protein Processing, Post-Translational / Q000235:genetics; D017293:Protein S / Q000737:chemistry / Q000302:isolation & purification*; D026403:S-Nitrosothiols / Q000737:chemistry; D015398:Signal Transduction / Q000235:genetics
chemical_list: D017293:Protein S; D026403:S-Nitrosothiols; D009569:Nitric Oxide; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D003545:Cysteine
keywords: Endothelial cell; Fluorescent probe; Imaging; Posttranslational modification; S-nitrosylation