title: Determination of muscle protein synthesis rates in fish using (2)H2O and (2)H NMR analysis of alanine.
abstract: Following administration of deuterated water ((2)H2O), the fractional synthetic rate (FSR) of a given endogenous protein can be estimated by (2)H-enrichment quantification of its alanine residues. Currently, this is measured by mass spectrometry following a derivatization procedure. Muscle FSR was measured by (1)H/(2)H NMR analysis of alanine from seabass kept for 6 days in 5% (2)H-enriched saltwater, following acid hydrolysis and amino acid isolation by cation-exchange chromatography of muscle tissue. The analysis is simple and robust, and provides precise measurements of excess alanine (2)H-enrichment in the 0.1-0.4% range from 50 mmol of alanine recovered from muscle protein.
authors.affiliation: Center for Neuroscience and Cell Biology, UC-BIOTEC, 3060-197, Cantanhede, Portugal.
authors.forename: Cátia
authors.initials: C
authors.lastname: Marques
authors.affiliation: Center for Neuroscience and Cell Biology, UC-BIOTEC, 3060-197, Cantanhede, Portugal.
authors.forename: Filipa
authors.initials: F
authors.lastname: Viegas
authors.affiliation: Center for Neuroscience and Cell Biology, UC-BIOTEC, 3060-197, Cantanhede, Portugal; Center for Functional Ecology, Department Life Sciences, University of Coimbra, 3000-456, Coimbra, Portugal.
authors.forename: João
authors.initials: J
authors.lastname: Rito
authors.affiliation: Center for Neuroscience and Cell Biology, UC-BIOTEC, 3060-197, Cantanhede, Portugal; Portuguese Diabetes Association (APDP), 1250-203, Lisboa, Portugal.
authors.forename: John
authors.initials: J
authors.lastname: Jones
authors.affiliation: Center for Neuroscience and Cell Biology, UC-BIOTEC, 3060-197, Cantanhede, Portugal; Center for Functional Ecology, Department Life Sciences, University of Coimbra, 3000-456, Coimbra, Portugal. Electronic address: iviegas@ci.uc.pt.
authors.forename: Ivan
authors.initials: I
authors.lastname: Viegas
mesh_terms: D000409:Alanine / Q000378:metabolism; D000818:Animals; D001492:Bass / Q000378:metabolism*; D017666:Deuterium Oxide / Q000737:chemistry*; D029941:Fish Proteins / Q000096:biosynthesis*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D014176:Protein Biosynthesis / Q000502:physiology*
chemical_list: D029941:Fish Proteins; D017666:Deuterium Oxide; D000409:Alanine
keywords: (2)H NMR; (2)H(2)O; Deuterated water; Fish protein synthesis; Fractional synthetic rate