title: Structure of proteins in eukaryotic compartments.
abstract: In-cell NMR in the yeast  Pichia pastoris  was used to study the influence of metabolic changes on protein structure and dynamics at atomic resolution. Induction of ubiquitin overexpression from the methanol induced AOX1 promoter results in the protein being localized in the cytosol and yields a well-resolved in-cell NMR spectrum. When  P. pastoris  is grown on a mixed carbon source containing both dextrose and methanol, ubiquitin is found in small storage vesicles distributed in the cytosol, and the resulting in-cell NMR spectrum is broadened. The sequestration of overexpressed proteins into storage vesicles, which are inaccessible to small molecules, was demonstrated for two unrelated proteins and two different strains of  P. pastoris , suggesting its general nature.
authors: Department of Chemistry, State University of New York at Albany, Albany, New York 12222, United States.
authors: Karl
authors: K
authors: Bertrand
authors: Sergey
authors: S
authors: Reverdatto
authors: David S
authors: DS
authors: Burz
authors: Richard
authors: R
authors: Zitomer
authors: Alexander
authors: A
authors: Shekhtman
mesh_terms: D000429:Alcohol Oxidoreductases / Q000235:genetics; D003001:Cloning, Molecular; D005947:Glucose / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D010843:Pichia / Q000166:cytology* / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D017510:Protein Folding; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis* / Q000235:genetics* / Q000378:metabolism; D025801:Ubiquitin / Q000032:analysis* / Q000235:genetics* / Q000378:metabolism; D015854:Up-Regulation; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D025801:Ubiquitin; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D001616:beta-Galactosidase; D005947:Glucose