PXD001210

PXD001210 is an original dataset announced via ProteomeXchange.

Title	An Enhanced in vivo SILAC model for Quantification of Drug Metabolism Enzymes
Description	Many of the enzymes involved in xenobiotic metabolism are maintained at a low basal level and are only synthesized in response to activation of upstream sensor/effector proteins. This induction can have implications in a variety of contexts, particularly during the study of the pharmacokinetics, pharmacodynamics and drug-drug interaction profile of a candidate therapeutic compound. Previously, we combined in vivo SILAC material with a targeted high resolution single ion monitoring (tHR/SIM) LC-MS/MS approach for quantification of 197 peptide pairs, representing 51 drug metabolism enzymes (DME), in mouse liver. However, as important enzymes (for example, cytochromes P450 (Cyp) of the 1a and 2b subfamilies) are maintained at low or undetectable levels in the liver of unstimulated metabolically-labelled mice, quantification of these proteins was unreliable. In the present study, we induced DME expression in labelled mice through synchronous ligand-mediated activation of multiple upstream nuclear receptors, thereby enhancing signals for proteins including Cyps 1a, 2a, 2b, 2c and 3a. With this enhancement, 115 unique, lysine-containing, Cyp-derived peptides were detected in the liver of a single animal, as opposed to 56 in a pooled sample from three uninduced animals. A total of 386 peptide pairs were quantified by tHR/SIM, representing 68 Phase I, 30 Phase II and 8 control proteins. This method was employed to quantify changes in DME expression in the hepatic cytochrome P450 reductase null (HRN) mouse. We observed compensatory induction of several enzymes, including Cyps 2b10, 2c29, 2c37, 2c54, 2c55, 2e1, 3a11 and 3a13, carboxylesterase (Ces) 2a and glutathione S-transferases (Gst) m2 and m3, along with down-regulation of hydroxysteroid dehydrogenases (Hsd) 11b1 and 17b6. Using DME-enhanced in vivo SILAC material with tHR/SIM, therefore, permits the robust analysis of multiple DME of import to xenobiotic metabolism, with improved utility for the study of drug pharmacokinetics, pharmacodynamics, and of chemically-treated and genetically-modified mouse models.
HostingRepository	PRIDE
AnnounceDate	2015-01-09
AnnouncementXML	Submission_2015-01-12_04:12:07.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD001210
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Kenneth MacLeod
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	Oxidation; Carbamidomethyl; Label:13C(6)
Instrument	LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2014-08-08 00:29:47	ID requested
1	2015-01-09 03:00:10	announced
⏵ 2	2015-01-12 04:12:08	announced	Updated publication reference for PubMed record(s): 25561501.

MacLeod AK, Fallon PG, Sharp S, Henderson CJ, Wolf CR, Huang JT, An enhanced in vivo stable isotope labeling by amino acids in cell culture (SILAC) model for quantification of drug metabolism enzymes. Mol Cell Proteomics, 14(3):750-60(2015) [pubmed]

curator keyword: Biomedical

submitter keyword: Drug metabolism, xenobiotic metabolism, drug-drug interaction, in vivo SILAC, pharmacology, toxicology, cytochrome P450

Jeffrey Tze-Jen Huang
contact affiliation	Jacqui Wood Cancer Centre, Medical Research Institute, Ninewells Hospital and Medical School, University of Dundee, James Arrott Drive, Dundee DD1 9SY, Scotland
contact email	j.t.j.huang@dundee.ac.uk
lab head
Kenneth MacLeod
contact affiliation	Department of Cancer Research
contact email	k.a.z.macleod@dundee.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD001210
     1. Label: PRIDE project
     2. Name: An Enhanced in vivo SILAC model for Quantification of Drug Metabolism Enzymes