title: High-yield expression of isotopically labeled peptides for use in NMR studies.
abstract: Fusion protein constructs of the 56 amino acid globular protein GB-1 with various peptide sequences, coupled with the incorporation of a histidine tag for affinity purification, have generated high-yield fusion protein constructs. Methionine residues were inserted into the constructs to generate pure peptides following CNBr cleavage, yielding a system that is efficient and cost effective for isotopic labeling of peptides for NMR studies and other disciplines such as mass spectroscopy. Six peptides of varying sequences and hydrophobicities were expressed using this GB-1 fusion protein technique and produced soluble fusion protein constructs in all cases. The ability to easily express and purify recombinant peptides in high yields is applicable for biomedical research and has medicinal and pharmaceutical applications.
authors.affiliation: Canadian Institutes of Health-Research Group in Protein Structure and Function, Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada T6G 2H7.
authors.forename: Darrin A
authors.initials: DA
authors.lastname: Lindhout
authors.forename: Angela
authors.initials: A
authors.lastname: Thiessen
authors.forename: Dean
authors.initials: D
authors.lastname: Schieve
authors.forename: Brian D
authors.initials: BD
authors.lastname: Sykes
mesh_terms: D000595:Amino Acid Sequence; D002247:Carbon Isotopes; D004926:Escherichia coli / Q000235:genetics; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D010452:Peptide Biosynthesis; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
chemical_list: D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D010455:Peptides; D011993:Recombinant Fusion Proteins