Dataset from proteomic analysis of human liver, lung, kidney and intestine microsomes

We provide detailed datasets from our analysis of proteins that are identified in human liver, lung, kidney and intestine microsomes by MS-based proteomics. Also included is a set of CYP450 enzymes and microsomal glutathione-S-transferase (MGSTs) activities in human liver microsomes. The data presented in this paper support the research article “Targeted label-free approach for quantification of epoxide hydrolase and glutathione transferases in microsomes” (Song et al., 2015) [1]. We expect that the data will contribute to the study of metabolism enzymes.


a b s t r a c t
We provide detailed datasets from our analysis of proteins that are identified in human liver, lung, kidney and intestine microsomes by MS-based proteomics. Also included is a set of CYP450 enzymes and microsomal glutathione-S-transferase (MGSTs) activities in human liver microsomes. The data presented in this paper support the research article "Targeted label-free approach for quantification of epoxide hydrolase and glutathione transferases in microsomes" (Song et al., 2015) [1]. We expect that the data will contribute to the study of metabolism enzymes. The proteins identified in human liver, lung, kidney and intestine microsomes with high sequence coverage.
Data from the LC-ESI-MS/MS analysis will provide researchers with detailed information on metabolism enzymes in human liver, lung, kidney and intestine microsomes.
Data from the enzyme activity analysis will enable researchers to observe the different activities of CYP450 enzymes and MGST enzyme in human liver microsomes.

Data
The data set shows the CYP450 enzymes and MGSTs activities in individual human liver microsomes (Table 1)

Mass spectrometric analysis
All of the samples were prepared following the previous study [1]. A reversed phase Waters nanoACQUITY column (1.7 µm, BEH130 C18, 100 µm i.d. × 100 µm, Waters Corp.) coupled to a Thermo LTQ Velos Orbitrap tandem mass spectrometer (Thermo Fisher Scientific) was used to identify the proteins in human liver, lung, kidney and intestine microsomes. Samples were eluted at 1.2 µL/min. t ¼ 0-5 min 99%A/1%B, t ¼ 5.1 min 85%A/15%B, t ¼ 50 min 40%A/60%B, t ¼ 55 min 15%A/85%B, t ¼ 55.1-65 min 99%A/1%B where A ¼ 97% water/3% acetonitrile/0.1% formic acid and B ¼ 0.1% formic acid in acetonitrile. Peptides were ionized via a nanoelectrospray ionization (ESI) source, and their mass spectra and collisionally induced dissociation (CID) fragmentation mass spectra were recorded. High resolution (60,000 resolving power), accurate mass spectra were recorded between m/z 395-2000 in~1.2 s on the orbitrap mass analyzer. While the next high-resolution mass spectrum was being acquired on the orbitrap, the LTQ Velos linear ion trap independently recorded CID fragmentation mass spectra of the 8 most abundant-S2 ions present in the previous orbitrap mass spectrum. During the course of a 60-min nano-LC-MS/MS run, this approach typically generated~3000 highresolution mass spectra and between 12,000-15,000 CID MS/MS spectra. Thermo-Finnegan Proteome Discoverer 2.0 software (Thermo Fisher Scientific) was used to interface with the Mascot (Matrix Science) protein database search engine. MS/MS spectral information was used by Mascot to search the SwissProt Protein database, and a decoy search was employed to establish a false discovery rate. MS Data processed using Mascot (S1, S2, S3, S4) are presented in the Supplementary information tables.
After preincubation at 37°C for 5 min, 10 μL of NADPH (20 mM) was added to initiate the reaction. After incubation, 400 μL acetonitrile containing internal standard was added to stop the reaction. The samples were centrifuged at 10,000 g for 10 min and the supernatants were concentrated by drying in vacuo. The residue was re-dissolved in 100 µL acetonitrile/water (50/50, v/v) and analyzed following the previous study [2,3].
Gutathione S-transferase activity was assayed following the method of Habig et al. [4] with 1 mM CDNB and 5 mM glutathione as the substrates.