Summary of Study ST001894
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001192. The data can be accessed directly via it's Project DOI: 10.21228/M8MX3X This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST001894 |
| Study Title | Involvement of Mieap in cardiolipin metabolism (part II) |
| Study Summary | Mass spectrometric data of Cardiolipin in Mice kidney (Mieap-WT vs. Mieap-KO), and Mice liver (Mieap-WT vs. Mieap-KO) |
| Institute | National Cancer Center Japan Research Institute |
| Last Name | Ikari |
| First Name | Naoki |
| Address | 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan |
| nikari@ncc.go.jp | |
| Phone | +81-3-3542-2511 |
| Submit Date | 2021-07-30 |
| Analysis Type Detail | LC-MS |
| Release Date | 2021-08-09 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001192 |
| Project DOI: | doi: 10.21228/M8MX3X |
| Project Title: | Mass Spectrometric Data of Cardiolipin in Cells under Mieap Expression |
| Project Summary: | Mass spectrometric data of cardiolipin in A549 (Ad-Mieap infected vs. non-infected) and LS174T cells (LS174T-cont vs. Mieap-KD) |
| Institute: | National Cancer Center Japan Research Institute |
| Last Name: | Ikari |
| First Name: | Naoki |
| Address: | 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan |
| Email: | nikari@ncc.go.jp |
| Phone: | +81-3-3542-2511 |
Subject:
| Subject ID: | SU001972 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL/6J Mieap-WT and Mieap-KO |
| Age Or Age Range: | 22-25 weeks of age |
| Gender: | Male |
| Animal Housing: | housed at 22 ± 2°C |
| Animal Light Cycle: | a 12 h light/dark cycle |
| Animal Feed: | free access to CE-2 (CLEA Japan) |
| Animal Water: | free access to water |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Genotype |
|---|---|---|
| SA175990 | Liver Mieap KO 1 | Mieap-knockout |
| SA175991 | Liver Mieap KO 2 | Mieap-knockout |
| SA175992 | Kidney Mieap KO 2 | Mieap-knockout |
| SA175993 | Kidney Mieap KO 1 | Mieap-knockout |
| SA175994 | Liver Mieap WT 2 | Wild-type |
| SA175995 | Liver Mieap WT 1 | Wild-type |
| SA175996 | Kidney Mieap WT 2 | Wild-type |
| SA175997 | Kidney Mieap WT 1 | Wild-type |
| Showing results 1 to 8 of 8 |
Collection:
| Collection ID: | CO001965 |
| Collection Summary: | The mouse tissues were snap-frozen in liquid nitrogen after weight measurement, and subsequently stored at -80°C until lipidomic analysis. |
| Sample Type: | Kidney and liver |
| Volumeoramount Collected: | 53.6-139.4mg |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR001984 |
| Treatment Summary: | The Mieap-knockout (Mieap-/-) mice were generated by using the Cre/loxP recombination system as previously reported[1]. Briefly, the floxed and trapped alleles were generated using a single construct bearing a gene-trap cassette doubly flanked by LoxP and FRT located between exons 5 and 8 of the mouse Mieap gene, which is located on chromosome 5. The Mieap homozygous (Mieap-/-) deficient mice were generated by mating breeding pairs of the Mieap heterozygous (Mieap+/-) mice. All mice were housed at 22 ± 2°C with a 12 h light/dark cycle with free access to food, CE-2 (CLEA Japan) and water. [1]Tsuneki, M., Nakamura, Y., Kinjo, T., Nakanishi, R., and Arakawa, H. (2015). Mieap suppresses murine intestinal tumor via its mitochondrial quality control. Sci. Rep. 5, 12472. |
| Treatment: | Knockout |
| Treatment Compound: | None |
| Animal Fasting: | None |
Sample Preparation:
| Sampleprep ID: | SP001978 |
| Sampleprep Summary: | Total lipids were extracted from samples using the Bligh-Dyer method. An aliquot of the organic phase was added to an equal volume of methanol before being loaded onto a DEAE-cellulose column (Wako Chemical) pre-equilibrated with chloroform. After successive washes with chloroform/methanol (1:1, v/v), acidic phospholipids were eluted with chloroform/methanol/HCl/water (12:12:1:1, v/v), followed by evaporation to dryness to yield a residue was soluble in methanol. |
| Extraction Method: | the Bligh-Dyer method |
Combined analysis:
| Analysis ID | AN003075 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | UltiMate 3000 (Thermo Fisher Scientific) |
| Column | Waters XBridge C18 (150 x 1.0mm,3.5um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap |
| Ion Mode | NEGATIVE |
| Units | pmol/mg |
Chromatography:
| Chromatography ID: | CH002275 |
| Chromatography Summary: | An aliquot of the lower/organic phase was added with an equal volume of methanol before being loaded onto a DEAE-cellulose column (Wako chemical) pre-equilibrated with chloroform. After successive washes with chloroform/methanol (1:1, v/v), the acidic phospholipids were eluted with chloroform/methanol/HCl/water (12:12:1:1, v/v), followed by evaporation to dryness to give a residue, which was resolved in methanol. |
| Instrument Name: | UltiMate 3000 (Thermo Fisher Scientific) |
| Column Name: | Waters XBridge C18 (150 x 1.0mm,3.5um) |
| Column Temperature: | 40 |
| Flow Gradient: | ratios of 60%/40% (0 min), 40%/60% (1 min), 20%/80% (9 min), 5%/95% (11-30 min), 95%/5% (31-35 min) and 60%/40% (45 min) |
| Flow Rate: | 25ul/min |
| Solvent A: | 50% isopropanol/10% methanol/40% water; 5 mM ammonium formate;0.05% ammonium hydroxide |
| Solvent B: | 100% isopropanol; 5 mM ammonium formate; 0.05% ammonium hydroxide |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002862 |
| Analysis ID: | AN003075 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Analyses were performed on a LC/MS/MS system consisting of a Q-Exactive Plus mass spectrometer (Thermo Fisher Scientific) equipped with an electrospray ionization source, and an UltiMate 3000 system (Thermo Fisher Scientific). The lipid samples were separated on Waters X-Bridge C18 column (3.5 μm, 150 mm × 1.0 mm i.d.) at 40°C using a gradient solvent system as follows: mobile phase A (isopropanol/methanol/water (5:1:4, v/v/v) supplemented with 5 mM ammonium formate and 0.05% ammonium hydroxide (28% in water))/mobile phase B (isopropanol supplemented with 5 mM ammonium formate and 0.05% ammonium hydroxide (28% in water)) ratios of 60%/40% (0 min), 40%/60% (1 min), 20%/80% (9 min), 5%/95% (11-30 min), 95%/5% (31-35 min) and 60%/40% (45 min). Flow rate was 25 μL/min. The source and ion transfer parameters applied were as followed: spray voltage 3.0 kV. For negative ionization modes, the sheath gas and the capillary temperature were maintained at 60 and 320 °C, respectively. The Orbitrap mass analyzer was operated at a resolving power of 70,000 in full-scan mode (scan range: 200–1800 m/z; automatic gain control (AGC) target:3e6) and of 35,000 in the Top 20 data-dependent MS2 mode (stepped normalized collision energy: 20, 30 and 40; isolation window: 4.0 m/z; AGC target: 1e5). Identification of CL molecular species was performed using with LipidSearch4.2 software (Mitsui knowledge industry). |
| Ion Mode: | NEGATIVE |
