title: A rapid method to attain isotope labeled small soluble peptides for NMR studies.
abstract: A widely applicable strategy is presented for efficient and rapid production of small water soluble peptides expressed as fusion proteins with the immunoglobulin-binding domain of streptococcal protein G. A simple extraction and purification scheme that includes a protease cleavage step to release the target peptide is described. The yield of authentic target peptide exceeds 10 mg per liter of culture. Production of U-13C, 15N and highly deuterated U-13C, 15N isotope labeled peptide is demonstrated for the 11 residue S2 peptide, corresponding to the C-terminus of the alpha-subunit of transducin, and the coiled coil trimerization domain from cartilage matrix protein (CMPcc), respectively. Heteronuclear two-dimensional NMR spectra are used for initial peptide characterization.
authors.affiliation: Structural Biology Institute, IBI-2, Research Centre Jülich, D-52425 Jülich, Germany. b.koenig@fz-juelich.de
authors.forename: Bernd W
authors.initials: BW
authors.lastname: Koenig
authors.forename: Marco
authors.initials: M
authors.lastname: Rogowski
authors.forename: John M
authors.initials: JM
authors.lastname: Louis
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D007553:Isotope Labeling / Q000379:methods*; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000032:analysis / Q000737:chemistry* / Q000235:genetics; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D012995:Solubility
chemical_list: D001426:Bacterial Proteins; C034284:IgG Fc-binding protein, Streptococcus; D010455:Peptides; D011993:Recombinant Fusion Proteins