title: A proteomics strategy for determining the synthesis and degradation rates of individual proteins in fish.
abstract: In order to study the protein dynamics in the tissues of fish we have developed a proteomics-based strategy to determine the rates of synthesis and degradation of individual proteins. We have demonstrated the feasibility of this approach by measuring the turnover of multiple isoforms of parvalbumin (β1-7) in the skeletal muscle of common carp (Cyprinus carpio). A stable isotope-labelled amino acid ([(2)H(7)] l-leucine) was administered to the carp via the diet and its incorporation into the isoforms of parvalbumin in muscle over time was monitored by LC-MS analysis of signature peptides. The relative isotope abundance was calculated and used to deconvolute the data. The β7 parvalbumin isoform had a rate of synthesis that was greater than the rate of degradation. In contrast the rate of degradation of the β5 isoform exceeded its rate of synthesis, whilst the analysis revealed that the other parvalbumin β-isoforms (β1, β2, β3, β4 and β6) had a rate of synthesis that was equal to the rate of degradation. This work has addressed a number of technical challenges and represents the first study to use proteomic approaches to measure the turnover of individual proteins in fish.
authors.affiliation: Department of Diabetes and Cardiovascular Science, University of the Highlands and Islands, Inverness, United Kingdom.
authors.forename: Mary K
authors.initials: MK
authors.lastname: Doherty
authors.forename: Philip
authors.initials: P
authors.lastname: Brownridge
authors.forename: Matthew A G
authors.initials: MA
authors.lastname: Owen
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Davies
authors.forename: Iain S
authors.initials: IS
authors.lastname: Young
authors.forename: Phillip D
authors.initials: PD
authors.lastname: Whitfield
mesh_terms: D000818:Animals; D002347:Carps / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000379:methods*; D008657:Metabolic Clearance Rate; D009124:Muscle Proteins / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D014018:Tissue Distribution
chemical_list: D009124:Muscle Proteins; D020543:Proteome