title: Stable isotope labeling of protein by Kluyveromyces lactis for NMR study.
abstract: Stable isotope labeling for proteins of interest is an important technique in structural analyses of proteins by NMR spectroscopy. Escherichia coli is one of the most useful protein expression systems for stable isotope labeling because of its high-level protein expression and low costs for isotope-labeling. However, for the expression of proteins with numerous disulfide-bonds and/or post-translational modifications, E. coli systems are not necessarily appropriate. Instead, eukaryotic cells, such as yeast Pichia pastoris, have great potential for successful production of these proteins. The hemiascomycete yeast Kluyveromyces lactis is superior to the methylotrophic yeast P. pastoris in some respects: simple and rapid transformation, good reproducibility of protein expression induction and easy scale-up of culture. In the present study, we established a protein expression system using K. lactis, which enabled the preparation of labeled proteins using glucose and ammonium chloride as a stable isotope source.
authors.affiliation: Japan Biological Informatics Consortium (JBiC), Aomi, Tokyo, 135-0064, Japan.
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Sugiki
authors.forename: Ichio
authors.initials: I
authors.lastname: Shimada
authors.forename: Hideo
authors.initials: H
authors.lastname: Takahashi
mesh_terms: D000643:Ammonium Chloride / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D007553:Isotope Labeling / Q000379:methods; D007716:Kluyveromyces / Q000235:genetics / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*
chemical_list: D005656:Fungal Proteins; D000643:Ammonium Chloride; D005947:Glucose