A proteomics approach to the study of absorption, distribution, metabolism, excretion, and toxicity.
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.
Nordvarg
Helena
H
GE Healthcare, Amersham Biosciences AB, Uppsala, Sweden.
Flensburg
John
J
Rönn
Ola
O
Ohman
Johan
J
Marouga
Rita
R
Lundgren
Bo
B
Haid
Daniel
D
Malmport
Eva
E
Goscinski
Jan
J
Hörnsten
Lena
L
Scigelova
Michaela
M
Bourin
Stephanie
S
Garberg
Per
P
Woffendin
Gary
G
Fenyö
David
D
Bergling
Hélène
H
Forsberg
Erik
E
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
D000178:Acrylamides; D020543:Proteome; C059413:Immobiline