Document Detail


A rapid method to attain isotope labeled small soluble peptides for NMR studies.
MedLine Citation:
PMID:  12766417     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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:
Bernd W Koenig; Marco Rogowski; John M Louis
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of biomolecular NMR     Volume:  26     ISSN:  0925-2738     ISO Abbreviation:  J. Biomol. NMR     Publication Date:  2003 Jul 
Date Detail:
Created Date:  2003-05-26     Completed Date:  2004-02-17     Revised Date:  2006-05-01    
Medline Journal Info:
Nlm Unique ID:  9110829     Medline TA:  J Biomol NMR     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  193-202     Citation Subset:  IM    
Affiliation:
Structural Biology Institute, IBI-2, Research Centre Jülich, D-52425 Jülich, Germany. b.koenig@fz-juelich.de
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MeSH Terms
Descriptor/Qualifier:
Animals
Bacterial Proteins / genetics,  metabolism
Gene Expression
Genetic Vectors / genetics
Isotope Labeling / methods*
Nuclear Magnetic Resonance, Biomolecular*
Peptides / analysis,  chemistry*,  genetics
Plasmids / genetics,  metabolism
Recombinant Fusion Proteins / chemistry,  metabolism
Solubility
Chemical
Reg. No./Substance:
0/Bacterial Proteins; 0/IgG Fc-binding protein, Streptococcus; 0/Peptides; 0/Recombinant Fusion Proteins

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