BioScience Trends. 2011;5(1):17-22. (DOI: 10.5582/bst.2011.v5.1.17)

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

Feng JH, Jing FB, Fang H, Gu LC, Xu WF


SUMMARY

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.


KEYWORDS: Histone deacetylase 8, S-nitrosylation, nitric oxide, S-nitrosoglutathione, S-nitrosocysteine

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