title: A solubility-enhancement tag (SET) for NMR studies of poorly behaving proteins.
abstract: Protein-fusion constructs have been used with great success for enhancing expression of soluble recombinant protein and as tags for affinity purification. Unfortunately the most popular tags, such as GST and MBP, are large, which hinders direct NMR studies of the fusion proteins. Cleavage of the fusion proteins often re-introduces problems with solubility and stability. Here we describe the use of N-terminally fused protein G (B1 domain) as a non-cleavable solubility-enhancement tag (SET) for structure determination of a dimeric protein complex. The SET enhances the solubility and stability of the fusion product dramatically while not interacting directly with the protein of interest. This approach can be used for structural characterization of poorly behaving protein systems, and would be especially useful for structural genomics studies.
authors: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
authors: P
authors: P
authors: Zhou
authors: A A
authors: AA
authors: Lugovskoy
authors: G
authors: G
authors: Wagner
mesh_terms: D000465:Algorithms; D000818:Animals; D051017:Apoptosis Regulatory Proteins; D001426:Bacterial Proteins / Q000737:chemistry*; D003851:Deoxyribonucleases / Q000737:chemistry*; D019281:Dimerization; D006801:Humans; D008970:Molecular Weight; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009994:Osmolar Concentration; D000075223:Poly-ADP-Ribose Binding Proteins; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D012995:Solubility
chemical_list: D051017:Apoptosis Regulatory Proteins; D001426:Bacterial Proteins; C034284:IgG Fc-binding protein, Streptococcus; D000075223:Poly-ADP-Ribose Binding Proteins; D011506:Proteins; D011993:Recombinant Fusion Proteins; C109664:caspase-activated DNase inhibitor; C113696:DFFB protein, human; D003851:Deoxyribonucleases