A rapid method to attain isotope labeled small soluble peptides for NMR studies.
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.
Koenig
Bernd W
BW
Structural Biology Institute, IBI-2, Research Centre Jülich, D-52425 Jülich, Germany. b.koenig@fz-juelich.de
Rogowski
Marco
M
Louis
John M
JM
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
D001426:Bacterial Proteins; C034284:IgG Fc-binding protein, Streptococcus; D010455:Peptides; D011993:Recombinant Fusion Proteins