title: Active-site-matched fluorescent probes for rapid and direct detection of vicinal-sulfydryl-containing peptides/proteins in living cells.
abstract: Vicinal-sulfydryl-containing peptides/proteins (VSPPs) play a crucial role in human pathologies. Fluorescent probes that are capable of detecting intracellular VSPPs in vivo would be useful tools to explore the mechanisms of some diseases. In this study, by regulating the spatial separation of two maleimide groups in a fluorescent dye to match that of two active cysteine residues contained in the conserved amino acid sequence (-CGPC-) of human thioredoxin, two active-site-matched fluorescent probes, o-Dm-Ac and m-Dm-Ac, were developed for real-time imaging of VSPPs in living cells. As a result, the two probes can rapidly respond to small peptide models and reduced proteins, such as WCGPCK (W-6), WCGGPCK (W-7), and WCGGGPCK (W-8), reduced bovine serum albumin (rBSA), and reduced thioredoxin (rTrx). Moreover, o-Dm-Ac displays a higher binding sensitivity with the above-mentioned peptides and proteins, especially with W-7 and rTrx. Furthermore, o-Dm-Ac was successfully used to rapidly and directly detect VSPPs both in vitro and in living cells. Thus, a novel probe-design strategy was proposed and the synthesized probe applied successfully in imaging of target proteins in situ.
authors.affiliation: College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Shandong Provincial Key Laboratory of Clean Production of Fine Chemicals, Shandong Normal University, Jinan, Shandong, 250014 (P.R. China); Department of Pharmaceutical Sciences, Binzhou Medical University, Yantai, Shandong, 264003 (P.R. China).
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Pan
authors.forename: Ziye
authors.initials: Z
authors.lastname: Liang
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.forename: Shanshan
authors.initials: S
authors.lastname: Wang
authors.forename: Fanpeng
authors.initials: F
authors.lastname: Kong
authors.forename: Kehua
authors.initials: K
authors.lastname: Xu
authors.forename: Bo
authors.initials: B
authors.lastname: Tang
mesh_terms: D000818:Animals; D002417:Cattle; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*
chemical_list: D005456:Fluorescent Dyes; D010455:Peptides; D011506:Proteins
keywords: fluorescent probes; imaging agents; peptides; proteins; thiols