Pub Date : 2017-08-31DOI: 10.1021/jacs.7b05725.s001
Weimin Xuan (1409371), Anzhi Yao (4402336), Peter G. Schultz (183240)
Sirtuins are NAD<sup>+</sup> dependent protein deacetylases, whichnare involved in many biological processes. We now report a novel geneticallynencoded fluorescent probe (EGFP-K85AcK) that responds to sirtuinsnin living cells. The probe design exploits a lysyl residue in EGFPnthat is essential for chromophore maturation, and is also an efficientndeacetylation substrate for sirtuins. Analysis of activity in Escherichia coli ΔcobB revealed that the probencan respond to various human sirtuins, including SIRT1, SIRT2, SIRT3nand SIRT5. We also directly monitored SIRT1 and SIRT2 activity innHEK293T cells with an mCherry fusion of EGFP-K85AcK, and showed thatnthis approach can be extended to other fluorescent proteins. Finally,nwe demonstrate that this approach can be used to examine the activitynof sirtuins toward additional lysyl posttranslational modifications,nand show that sirtuins can act as erasers of HibK modified proteins.
{"title":"GeneticallynEncoded Fluorescent Probe for DetectingnSirtuins in Living Cells","authors":"Weimin Xuan (1409371), Anzhi Yao (4402336), Peter G. Schultz (183240)","doi":"10.1021/jacs.7b05725.s001","DOIUrl":"https://doi.org/10.1021/jacs.7b05725.s001","url":null,"abstract":"Sirtuins are NAD<sup>+</sup> dependent protein deacetylases, whichnare involved in many biological processes. We now report a novel geneticallynencoded fluorescent probe (EGFP-K85AcK) that responds to sirtuinsnin living cells. The probe design exploits a lysyl residue in EGFPnthat is essential for chromophore maturation, and is also an efficientndeacetylation substrate for sirtuins. Analysis of activity in Escherichia coli ΔcobB revealed that the probencan respond to various human sirtuins, including SIRT1, SIRT2, SIRT3nand SIRT5. We also directly monitored SIRT1 and SIRT2 activity innHEK293T cells with an mCherry fusion of EGFP-K85AcK, and showed thatnthis approach can be extended to other fluorescent proteins. Finally,nwe demonstrate that this approach can be used to examine the activitynof sirtuins toward additional lysyl posttranslational modifications,nand show that sirtuins can act as erasers of HibK modified proteins.","PeriodicalId":522845,"journal":{"name":"Figshare","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2017-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147904562","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：