PXD029639

PXD029639 is an original dataset announced via ProteomeXchange.

Title	Amino acid and heavy water labelling to obtain protein turnover rates in intact adult animals
Description	Both labelling studies were conducted with C57/BL6J adult male mice, derived from the same supplier and maintained under identical conditions. Eleven adult male C57BL/6JOlaHsd mice (Harlan UK Ltd., Shardlow, UK) of over 12 months of age at the start of the experiment were housed individually in 48 x 15 x 15 cm polypropylene cages (NKP Cages Ltd., Coalville, UK). Each cage contained substrate (Corn Cob Absorb 10-14 substrate), paper wool nest material and environmental enrichment. Food (LabDiet 5002 Certified Rodent Diet, Purina Mills, St. Louis, USA) and water were provided ad libitum. The mice were maintained on a reversed photo-period (light, 12 h; dark, 12 h; lights on at 2000 hrs) and at 19-21 oC ± 2 oC. Standard laboratory diet was replaced with a semisynthetic diet with the inclusion of [13C6]lysine at a relative isotope abundance (RIA) of 0.5. The dietary pellets were dissociated with water containing the dissolved [13C6]lysine to form a thick paste and mixed extensively. Once homogeneous, the paste was then extruded into strips 1 cm across and dried in a commercial foodstuff drying oven at 40 oC. The mice had access to the labelled diet for varying amounts of time: 0, 1, 2, 3, 4, 6, 9, 12, 17, 22 or 30 days. The day that the animals were introduced to the labelled diet was staggered for all culls and dissections to take place on the same day. All mice were humanely killed on day 30 and dissected to recover liver, kidney, heart and pooled hindlimb skeletal muscle from each animal. All resected tissue was frozen at -80 oC prior to analysis. For the heavy water labelling protocol, all animals were provided free access to LabDiet 5002. At the start of the experiment, mice were injected with two successive 0.5 mL injections, 4 hours apart, of 0.15 M sodium chloride dissolved in deuterated water. Thereafter, mice were given free access to 8% (v/v) [2H2]O for the duration of the experiment. After 0, 1, 2, 3, 6, 7, 9, 13, 16, 21, 24 and 30 days, mice were killed and dissected exactly as described for the amino acid labelling experiment and tissues were stored at -80 oC prior to analysis.
HostingRepository	jPOST
AnnounceDate	2021-12-13
AnnouncementXML	Submission_2023-12-18_08:59:35.012.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Edward Lau
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	S-carboxamidomethyl-L-cysteine; L-methionine sulfoxide; alpha-amino acetylated residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2021-11-09 14:41:21	ID requested
1	2021-12-13 08:53:43	announced
2	2022-09-18 03:31:54	announced	2022-09-18: Updated FTP location.
⏵ 3	2023-12-18 08:59:35	announced	2023-12-19: Updated PubMed.

Hammond DE, Simpson DM, Franco C, Wright Muelas M, Waters J, Ludwig RW, Prescott MC, Hurst JL, Beynon RJ, Lau E, Harmonizing Labeling and Analytical Strategies to Obtain Protein Turnover Rates in Intact Adult Animals. Mol Cell Proteomics, 21(7):100252(2022) [pubmed]

submitter keyword: mouse, heavy water, silac, turnover

Robert J Beynon
lab head
Edward Lau
contact affiliation	University of Colorado
dataset submitter

jPOST dataset URI

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.jpostdb.org/JPST001378/