Expression, purification, and S-nitrosylation of recombinant histone deacetylase 8 in Escherichia coli

• Published in: BioScience Trends Vol. 5; no. 1; pp. 17 - 22
• Main Authors: Feng, Jinhong, Jing, Fanbo, Fang, Hao, Gu, Lichuan, Xu, Wenfang
• Format: Journal Article
• Language: English
• Published: Japan International Research and Cooperation Association for Bio & Socio-Sciences Advancement 01.01.2011
• Subjects: Blotting, Western; Cysteine - analogs & derivatives; Cysteine - chemical synthesis; Cysteine - metabolism; DNA Primers - genetics; Electrophoresis, Polyacrylamide Gel; Escherichia coli; Histone deacetylase 8; Histone Deacetylases - isolation & purification; Histone Deacetylases - metabolism; Humans; In Vitro Techniques; nitric oxide; Nitric Oxide - metabolism; Plasmids - genetics; Recombinant Proteins - metabolism; Repressor Proteins - isolation & purification; Repressor Proteins - metabolism; S-nitrosocysteine; S-nitrosoglutathione; S-Nitrosoglutathione - chemical synthesis; S-Nitrosoglutathione - metabolism; S-Nitrosothiols - chemical synthesis; S-Nitrosothiols - metabolism; S-nitrosylation
• ISSN: 1881-7815; 1881-7823
• DOI: 10.5582/bst.2011.v5.1.17

Histone deacetylase (HDAC) 8 is a zinc ion dependent enzyme involved in removing the acetyl group from the core histones and other proteins which belong to Class I HDACs. It was reported that nitric oxide (NO) is a key regulator of HDAC function and S-nitrosylation of HDAC2 induces chromatin remodelling in neurons. This work reports the successful recombinant expression of human HDAC8 in Escherichia coli with two plasmids and the purification and S-nitrosylation in vitro. It was found that HDAC8 can be S-nitrosylated by the NO donor S-nitrosoglutathione (GSNO) in vitro, and the activity of HDAC8 was significantly inhibited when incubated with GSNO and S-nitrosocysteine in a time- and dosage-dependent manner, but sodium nitroprusside (SNP), and dithiothreitol cannot reverse this inhibition. These observations support and extend the concept that NO may regulate HDAC8 function by S-nitrosylation.