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High‐yield expression of isotopically labeled peptides for use in NMR studies
Author(s) -
Lindhout Darrin A.,
Thiessen Angela,
Schieve Dean,
Sykes Brian D.
Publication year - 2003
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.0376003
Subject(s) - peptide , histidine , chemistry , nuclear magnetic resonance spectroscopy , yield (engineering) , recombinant dna , biochemistry , fusion protein , amino acid , cleavage (geology) , peptide sequence , lysine , mass spectrometry , combinatorial chemistry , chromatography , stereochemistry , biology , materials science , paleontology , fracture (geology) , metallurgy , gene
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.