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Protocol to purify the histone deacetylase SIRT6 and assess its activity in vitro.
Tian, Yuan; Feng, Tingting; Hou, Tianyun; Zhu, Wei-Guo.
Affiliation
  • Tian Y; Guangdong Key Laboratory of Genome Stability and Human Disease Prevention, Shenzhen University International Cancer Center, Marshall Laboratory of Biomedical Engineering, Department of Biochemistry and Molecular Biology, Shenzhen University School of Medicine, Shenzhen 518055, China.
  • Feng T; Guangdong Key Laboratory of Genome Stability and Human Disease Prevention, Shenzhen University International Cancer Center, Marshall Laboratory of Biomedical Engineering, Department of Biochemistry and Molecular Biology, Shenzhen University School of Medicine, Shenzhen 518055, China.
  • Hou T; Guangdong Key Laboratory of Genome Stability and Human Disease Prevention, Shenzhen University International Cancer Center, Marshall Laboratory of Biomedical Engineering, Department of Biochemistry and Molecular Biology, Shenzhen University School of Medicine, Shenzhen 518055, China.
  • Zhu WG; Guangdong Key Laboratory of Genome Stability and Human Disease Prevention, Shenzhen University International Cancer Center, Marshall Laboratory of Biomedical Engineering, Department of Biochemistry and Molecular Biology, Shenzhen University School of Medicine, Shenzhen 518055, China. Electronic addr
STAR Protoc ; 4(2): 102206, 2023 Mar 29.
Article in En | MEDLINE | ID: mdl-36995934
ABSTRACT
The histone deacetylase known as sirtuin 6 (SIRT6) deacetylates both histone and non-histone proteins but has low deacetylase activity in vitro. Here, we present a protocol to monitor SIRT6-mediated deacetylation of long-chain acyl-CoA synthase 5 in the presence of palmitic acid. We describe the purification of His-SIRT6 and a Flag-tagged substrate. We then detail a deacetylation assay protocol that can be widely applied to study other SIRT6-mediated deacetylation events and the effect of SIRT6 mutations on its activity. For complete details on the use and execution of this protocol, please refer to Hou et al. (2022).1.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Language: En Journal: STAR Protoc Year: 2023 Document type: Article Affiliation country: China
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Full text: 1 Collection: 01-internacional Database: MEDLINE Language: En Journal: STAR Protoc Year: 2023 Document type: Article Affiliation country: China