PXD054543-1

PXD054543 is an original dataset announced via ProteomeXchange.

Title	Northern elephant seal juvenile plasma proteome response to repeated ACTH administration
Description	Plasma proteome response to repeated ACTH administration in juvenile northern elephant seals. Five ul of EDTA plasma were denatured with 200 ul of 5 percent w/v SDC and 5 mM TCEP in 50 mM AmBiC at 60C for 1 hour, followed by alkylation with 20 mM CAA for 30 minutes in the dark at room temp. Samples were diluted to 0.5 percent SDC and proteins were digested using trypsin at a 1 to 50 ug enzyme to protein ratio for 16 hours at 37C. TFA was added to a final concentration of 1.0 percent to precipitate SDC, which was removed by centrifugation and extraction of the supernatant. Digested peptides were lyophilized, resuspended in 0.1 percent TFA, and desalted using Pierce Peptide Desalting Spin Columns. Eluted peptides were lyophilized and resuspended in 0.1 percent FA in LC-MS grade water. Three injections were used for each sample. For each run, 5 uL of the sample (1 ug total) was loop injected onto a reversed-phase trap column (Acclaim PepMap 100 C18 LC column; 75 um i.d. x 2 cm, 3 um particle size, 100 A pore size, Thermo Fisher Scientific, USA) by a Dionex Ultimate 3000 autosampler. Peptides were eluted onto a reversed-phase analytical column set at 35C for HPLC (EASY-SprayTM C18 LC column; 75 um i.d. x 15 cm, 100 A, Thermo Fisher Scientific, USA). Solvent A was water and B was acetonitrile (ACN), respectively (both with 0.1 percent formic acid). During each chromatographic run, which lasted 140 min, flow rates were held at 300 nl/min. Solvent B (ACN) was used as follows: 2 percent at 5 min, 2-22 percent at 75 min, 22-38 percent at 100 min, 38-95 percent at 105 min, 95 percent returning to 2 percent at 115 min, and lastly maintained at 2 percent at 140 min. Mass spectrometry analysis was performed using Orbitrap Fusion Tribrid mass spectrometer equipped with an EASY-SprayTM ion source and operated by Xcalibur 4.0 software. MS1 spectra were resolved by the orbitrap with a resolution of 120,000, scan range of 200-1400 m/z, RF lens of 60 percent, AGC target of 1.0e6, and max injection time of 50 ms. Precursor ions were isolated by quadrupole and fragmented using HCD with a collision energy of 28 percent. MS2 product ions were resolved by the orbitrap (resolution: 30,000, AGC target: 5.0e5, first mass: 100 m/z, and max injection time: 150 ms).
HostingRepository	MassIVE
AnnounceDate	2024-08-02
AnnouncementXML	Submission_2024-08-02_12:20:36.103.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Jane Khudyakov
SpeciesList	scientific name: Mirounga angustirostris; common name: Northern elephant seal; NCBI TaxID: 9716;
ModificationList	Carbamidomethyl; Acetyl; Oxidation
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2024-08-02 09:53:16	ID requested
⏵ 1	2024-08-02 12:20:36	announced

no publication

submitter keyword: plasma, stress, seal

Jane Khudyakov
contact affiliation	University of the Pacific
contact email	jkhudyakov@pacific.edu
lab head
Jane Khudyakov
contact affiliation	University of the Pacific
contact email	jkhudyakov@pacific.edu
dataset submitter

MassIVE dataset URI

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