title: A proteomics approach to the study of absorption, distribution, metabolism, excretion, and toxicity.
abstract: A proteomics approach was used to identify liver proteins that displayed altered levels in mice following treatment with a candidate drug. Samples from livers of mice treated with candidate drug or untreated were prepared, quantified, labeled with CyDye DIGE Fluors, and subjected to two-dimensional electrophoresis. Following scanning and imaging of gels from three different isoelectric focusing intervals (3-10, 7-11, 6.2-7.5), automated spot handling was performed on a large number of gel spots including those found to differ more than 20% between the treated and untreated condition. Subsequently, differentially regulated proteins were subjected to a three-step approach of mass spectrometry using (a) matrix-assisted laser desorption/ionization time-of-flight mass spectrometry peptide mass fingerprinting, (b) post-source decay utilizing chemically assisted fragmentation, and (c) liquid chromatography-tandem mass spectrometry. Using this approach we have so far resolved 121 differentially regulated proteins following treatment of mice with the candidate drug and identified 110 of these using mass spectrometry. Such data can potentially give improved molecular insight into the metabolism of drugs as well as the proteins involved in potential toxicity following the treatment. The differentially regulated proteins could be used as targets for metabolic studies or as markers for toxicity.
authors.affiliation: GE Healthcare, Amersham Biosciences AB, Uppsala, Sweden.
authors.forename: Helena
authors.initials: H
authors.lastname: Nordvarg
authors.forename: John
authors.initials: J
authors.lastname: Flensburg
authors.forename: Ola
authors.initials: O
authors.lastname: Rönn
authors.forename: Johan
authors.initials: J
authors.lastname: Ohman
authors.forename: Rita
authors.initials: R
authors.lastname: Marouga
authors.forename: Bo
authors.initials: B
authors.lastname: Lundgren
authors.forename: Daniel
authors.initials: D
authors.lastname: Haid
authors.forename: Eva
authors.initials: E
authors.lastname: Malmport
authors.forename: Jan
authors.initials: J
authors.lastname: Goscinski
authors.forename: Lena
authors.initials: L
authors.lastname: Hörnsten
authors.forename: Michaela
authors.initials: M
authors.lastname: Scigelova
authors.forename: Stephanie
authors.initials: S
authors.lastname: Bourin
authors.forename: Per
authors.initials: P
authors.lastname: Garberg
authors.forename: Gary
authors.initials: G
authors.lastname: Woffendin
authors.forename: David
authors.initials: D
authors.lastname: Fenyö
authors.forename: Hélène
authors.initials: H
authors.lastname: Bergling
authors.forename: Erik
authors.initials: E
authors.lastname: Forsberg
mesh_terms: D000178:Acrylamides; D000818:Animals; D002623:Chemistry Techniques, Analytical; D015180:Electrophoresis, Gel, Two-Dimensional; D051379:Mice; D020543:Proteome / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000178:Acrylamides; D020543:Proteome; C059413:Immobiline