title: High-level bacterial expression and purification of human SirT2 protein for NMR studies.
abstract: Silent information regulator 2 (Sir2) proteins are a class of protein deacetylase enzymes that play key roles in transcriptional gene silencing, DNA repair, and aging. Here, we describe the high-level bacterial expression and purification of a human SirT2 construct that yields high resolution NMR spectra. By removing the N-terminal helix alpha0 and using Thioredoxin as a fusion partner, greater than 10 mg/L of purified protein can be obtained from minimal media. The protein is fully functional and enables NMR-based screening and structural studies of this important protein.
authors.affiliation: Global Pharmaceutical Discovery Division, Abbott Laboratories, Abbott Park, IL, USA.
authors.forename: Chaohong
authors.initials: C
authors.lastname: Sun
authors.forename: Danying
authors.initials: D
authors.lastname: Song
authors.forename: Patrick A
authors.initials: PA
authors.lastname: Marcotte
authors.forename: Paul L
authors.initials: PL
authors.lastname: Richardson
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Hajduk
mesh_terms: D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D019906:Nuclear Magnetic Resonance, Biomolecular; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*; D056565:Sirtuin 2; D037761:Sirtuins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D013879:Thioredoxins / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D013879:Thioredoxins; C474199:SIRT2 protein, human; D056565:Sirtuin 2; D037761:Sirtuins