c terminal his tag  (Sino Biological)

Journal: RSC Medicinal Chemistry

Article Title: Structure–activity relationships of hydrophobic small molecule irreversible inhibitors of tissue transglutaminase

doi: 10.1039/d5md00815h

Figure Lengend Snippet: Conformational equilibrium and inhibition of TG2. A) Closed conformation (based on PDB code 4PYG) with assembled GTP-binding site shown in magenta. B) Open conformation (based on PDB code 2Q3Z) with substrate binding site shown in blue. C) Reaction with an irreversible inhibitor (cyan) locks TG2 in the open conformation.

Article Snippet: The enzyme was expressed and purified according to a modified procedure developed by Han et al. A swab of Novagen Rosetta 2 (DE3) E. coli cells, which had been transformed with a plasmid containing the C-terminal His-tag TG2 gene (purchased from Addgene, plasmid #100719), was added to two test tubes with 5 mL of sterile Terrific Broth (TB) medium and 5 μL each of kanamycin (50 mg mL −1 ) and chloramphenicol (34 mg mL −1 ) stock solutions.

Techniques: Inhibition, Binding Assay