PXD000675

PXD000675 is an original dataset announced via ProteomeXchange.

Title	A Targeted in Vivo SILAC Approach for Quantification of Drug Metabolism Enzymes: Regulation by the Constitutive Androstane Receptor
Description	The modulation of drug metabolism enzyme (DME) expression by therapeutic agents is a central mechanism of drug–drug interaction and should be assessed as early as possible in preclinical drug development. Direct measurement of DME levels is typically achieved by Western blotting, qPCR, or microarray, but these techniques have their limitations; antibody cross-reactivity among highly homologous subfamilies creates ambiguity, while discordance between mRNA and protein expression undermines observations. The aim of this study was to design a simple targeted workflow by combining in vivo SILAC and label-free proteomics approaches for quantification of DMEs in mouse liver, facilitating a rapid and comprehensive evaluation of metabolic potential at the protein level. A total of 197 peptides, representing 51 Phase I and Phase II DMEs, were quantified by LC-MS/MS using targeted high resolution single ion monitoring (tHR/SIM) with a defined mass-to-charge and retention time window for each peptide. In a constitutive androstane receptor (Car) activated mouse model, comparison of tHR/SIM-in vivo SILAC with Western blotting for analysis of the expression of cytochromes P450 was favorable, with agreement in fold-change values between methods. The tHR/SIM-in vivo SILAC approach therefore permits the robust analysis of multiple DME in a single protein sample, with clear utility for the assessment of the drug–drug interaction potential of candidate therapeutic compounds.
HostingRepository	PRIDE
AnnounceDate	2014-07-24
AnnouncementXML	Submission_2014-07-24_03:48:41.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Kenneth MacLeod
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2014-01-08 03:23:48	ID requested
1	2014-02-14 02:54:10	announced
⏵ 2	2014-07-24 03:48:42	announced	Updated project metadata.

Macleod AK, Zang T, Riches Z, Henderson CJ, Wolf CR, Huang JT, A targeted in vivo SILAC approach for quantification of drug metabolism enzymes: regulation by the constitutive androstane receptor. J Proteome Res, 13(2):866-74(2014) [pubmed]

curator keyword: Biomedical

submitter keyword: drug metabolism

drug−drug interaction

constitutive androstane receptor

protein quantification

targeted in vivo SILAC

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. PXD000675
     1. Label: PRIDE project
     2. Name: A Targeted in Vivo SILAC Approach for Quantification of Drug Metabolism Enzymes: Regulation by the Constitutive Androstane Receptor