title: A FRET-based assay for screening SIRT5 specific modulators.
abstract: A fluorogenic assay for SIRT5 has been developed to screen their small molecule modulators based on the recent discovery that SIRT5 is a demalonylase and desuccinylase. However, this assay uses a fluorogenic peptide containing 7-amino-4-methylcoumarin (AMC), which becomes the cause of false positive hits from the screening. To overcome this, we have developed an alternative method called a FRET-based assay, which will be reliable and useful for screening SIRT5 modulators in a high-throughput format since no AMC group present in this assay.
authors.affiliation: College of Basic Medical Science, Guiyang Medical University, Guiyang, Guizhou 550004, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: College of Pharmacy, Guiyang Medical University, Guiyang, Guizhou 550004, China; Engineering Research Center for the Development and Application of Ethnic Medicine and TCM, Ministry of Education, Guiyang Medical University, Guiyang, Guizhou 550004, China.
authors.forename: Wenfei
authors.initials: W
authors.lastname: Huang
authors.affiliation: College of Pharmacy, Guiyang Medical University, Guiyang, Guizhou 550004, China; Engineering Research Center for the Development and Application of Ethnic Medicine and TCM, Ministry of Education, Guiyang Medical University, Guiyang, Guizhou 550004, China.
authors.forename: Ling
authors.initials: L
authors.lastname: You
authors.affiliation: College of Pharmacy, Guiyang Medical University, Guiyang, Guizhou 550004, China; Engineering Research Center for the Development and Application of Ethnic Medicine and TCM, Ministry of Education, Guiyang Medical University, Guiyang, Guizhou 550004, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Xie
authors.affiliation: College of Pharmacy, Guiyang Medical University, Guiyang, Guizhou 550004, China; Engineering Research Center for the Development and Application of Ethnic Medicine and TCM, Ministry of Education, Guiyang Medical University, Guiyang, Guizhou 550004, China. Electronic address: binhe@gmc.edu.cn.
authors.forename: Bin
authors.initials: B
authors.lastname: He
mesh_terms: D000595:Amino Acid Sequence; D003374:Coumarins / Q000737:chemistry; D057075:Enzyme Assays; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D007700:Kinetics; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D037761:Sirtuins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013379:Substrate Specificity; D004124:p-Dimethylaminoazobenzene / Q000031:analogs & derivatives / Q000737:chemistry
chemical_list: D003374:Coumarins; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D010455:Peptides; C063127:4-(4-dimethylaminophenylazo)benzoic acid; D004124:p-Dimethylaminoazobenzene; D037761:Sirtuins; C028743:7-amino-4-methylcoumarin
keywords: Deacetylation; Desuccinylation; FRET assay; Fluorogenic assay; Sirtuin