PXD052641-1

PXD052641 is an original dataset announced via ProteomeXchange.

Title	Proteomic profiles of adult, aged, and hypertrophic mouse hearts
Description	All animal protocols were approved by the Institutional Animal Care and Use Committee (IACUC) guidelines at the University of Colorado Denver/Anschutz Medical Campus. For normal and hypertrophic mouse hearts, wild-type C57BL/6J mice were purchased from Jackson Laboratories (Bar Harbor, ME, USA) and housed in a temperature-controlled environment on a 12-h light/dark cycle and with free access to normal chow diet and water under National Institutes of Health (NIH) guidelines for the Care and Use of Laboratory Animals. Adult (16-17 wk) and aged (86 wk) mice were administered with saline or 20 mg/kg/d isoproterenol by micro-osmotic pump (ALZET) implantation for a duration of 7 or 14 days (male and female, n = 2 per sex per age group, 32 animals total). The animals were euthanized, followed by measurement of body weight, heart weight and tibia length. Left ventricles were collected and kept frozen at –80 °C. Tissue homogenization was performed with a handheld homogenizer (OMNI International). Samples were placed on ice for 1 minute to allow cooling down. Repeat for a total of 3 rounds of homogenization. Homogenized tissues were sonicated with a handheld sonicator (Fisher Model 120), at 40% amplitude for 1 s, pause for 5 s, for 15 cycles. The samples were then centrifuged at 5,000 × g for 1 min at 4°C, and vortex for 10 s, and repeated for three rounds of sonication and centrifugation. The extracted protein samples were further centrifuged at 14,000 × g for 15 min at 4°C to collect supernatants and measure protein concentration with the BCA protein assay kit following manufacturer instructions. The samples were then reduced, alkylated, and digested with sequencing-grade trypsin. The digested peptides were labeled with TMT 10-plex isobaric tags (Thermo), and fractionated using high-pH reversed-phase columns (Pierce) following manufacturer instructions. Mass spectrometry was performed on a Q-Exactive HF orbitrap high-resolution mass spectrometer (Thermo) coupled to a Thermo Easy-nLC UPLC liquid chromatography system. The LC gradient was as follows: 0 – 105 minutes, 0 – 30% B; 105 – 110 minutes, 30 – 70% B, 110 – 115 minutes: 70 – 100% B; 115 – 120 minutes: hold; at 300 nL/min. Mass spectrometry data were acquired in data dependent acquisition (DDA) mode using typical settings including 60,000 MS1 and MS2 resolution, 3e6 MS1 AGC; 2e5 MS2 AGC; 110 ms maximum IT; 15 TopN; isolation window 0.7 m/z; normalized CE 28, 30, 32; and 30 seconds dynamic exclusion. Mass spectrometry data was searched using Sage v.0.14.5 aarch64-apple-darwin build to identify proteins and to measure peptide MS2 TMT and MS1 LFQ (label-free quantification) intensity, using typical settings including: missed_cleavages: 2, precursor_tol: –20 to +20 ppm; fragment_tol: –20 to +20 ppm; isotope: false; static_mods: C 57.0215 K 229.1629; variable mods: M 15.9949 ^ 229.1629 S 229.1629; generate_decoys: true; isotope_errors: 0 – 3; quant: tmt: Tmt10; quant: lfq: true, The database used was retrieved on 2023-09-15 from UniProt to contain 25,530 Swiss-Prot Mus musculus reviewed canonical and isoform entries. A peptide-level multiple testing adjusted q value cutoff of 0.01 was used for confident identification.
HostingRepository	jPOST
AnnounceDate	2025-07-23
AnnouncementXML	Submission_2025-07-23_14:08:24.921.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Edward Lau
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	TMT; L-methionine sulfoxide; TMT; S-carboxamidomethyl-L-cysteine
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2024-05-28 14:14:34	ID requested
⏵ 1	2025-07-23 14:08:25	announced

Dataset with its publication pending

submitter keyword: mouse, heart, aging, hypertrophy, heart failure

Maggie Lam, PhD
lab head
Edward Lau
contact affiliation	University of Colorado
dataset submitter

jPOST dataset URI

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