PXD009622
PXD009622 is an original dataset announced via ProteomeXchange.
Dataset Summary
Title | Quantitative Proteomics Reveals Antibiotics Resistance Function of Outer Membrane Proteins in Aeromonas hydrophila |
Description | Each colony of A. hydrophila and the OXY-R strain were incubated in 5 mL LB medium overnight, and then diluted in 100 mL fresh LB medium at a ratio of 1:100 and subsequently cultured until the optical density at 600 nm (OD600) reached 1.0. The cultures were harvested via centrifuging for 20 min at 10,000 x g, 4°C, and then the bacterial cells were washed with cold phosphate buffered saline (PBS, pH 7.4) for three times. The cell pellets were resuspending in the 10 mL ultrasonic buffer (50 mM Tris-HCl, pH 7.4, 1 mM PMSF) and disrupted with intermittent sonic oscillation for a total of 30 minutes at 9 seconds intervals on ice. Subsequently, the cell debris and unbroken cells were separated by centrifugation at 8,000 x g for 20 min at 4°C. Then the supernatant was centrifuged at 100,000 x g for 1 h at 4°C in the Optima LE-80 K Ultracentrifuge (Beckman, Palo Alto, CA, USA). After the pellet was dissolved with 2% sodium lauroyl sarcosine (in 50 mM Tris-HCl, pH 7.5) for 40 min at room temperature (RT), the pellets were ultracentrifuged again at 100,000 x g for 1 h at 4°C. Finally, the precipitate was dissolved in an appropriate volume of SDT buffer (4 % SDS, 0.1 M DTT [dithiothreitol], and 0.5 M triethylammonium bicarbonate buffer [TEAB, pH 8.5]). The protein concentration was determined using the Bradford method and then stored at -20°C until subsequent use.The proteins were digested with trypsin after being reduced with DTT and alkylated with Iodoacetamide using a filter-aided sample preparation method (Tanca et al., 2013; Li et al., 2016b). After washing three times with 0.5 M TEAB followed by fractionation using the 10 kDa ultrafiltration system (Millipore, Billerica, MA, USA), about 100 μg digested peptide from each group, including two biological replicates, was taken out for further labeled using TMT isobaric and isotopic mass-tagging kits (Thermo Fisher Scientific, MA, USA), which was performed according to instructions from the kit. |
HostingRepository | PRIDE |
AnnounceDate | 2018-11-23 |
AnnouncementXML | Submission_2018-11-23_06:55:02.xml |
DigitalObjectIdentifier | https://dx.doi.org/10.6019/PXD009622 |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Supported dataset by repository |
PrimarySubmitter | Xiangmin Lin |
SpeciesList | scientific name: Aeromonas hydrophila subsp. hydrophila ATCC 7966; NCBI TaxID: 380703; |
ModificationList | Ammonia-loss; Deamidated; TMT6plex; Dehydrated; Oxidation; Carbamidomethyl |
Instrument | TripleTOF 5600 |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
---|---|---|---|
0 | 2018-04-27 02:18:19 | ID requested | |
⏵ 1 | 2018-11-23 06:55:03 | announced |
Publication List
Yao Z, Sun L, Wang Y, Lin L, Guo Z, Li D, Lin W, Lin X, . Front Cell Infect Microbiol, 8():390(2018) [pubmed] |
Keyword List
curator keyword: Biomedical |
submitter keyword: Aeromonas hydrophila, Outer Membrane Proteins, Antibiotics resistance, LC-MS/MS |
Contact List
Xiangmin Lin | |
---|---|
contact affiliation | College of Life Sciences, Fujian Agriculture and Forestry University |
contact email | xiangmin@fafu.edu.cn |
lab head | |
Xiangmin Lin | |
contact affiliation | Fujian Agricultural and Forestry University |
contact email | xiangmin@fafu.edu.cn |
dataset submitter |
Full Dataset Link List
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PRIDE project URI |
Repository Record List
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