title: Proteomic evaluation of myofibrillar carbonylation in chilled fish mince and its inhibition by catechin.
abstract: The present study investigates the susceptibility of individual myofibrillar proteins from mackerel (Scomber scombrus) mince to undergo carbonylation reactions during chilled storage, and the antioxidant capacity of (+)-catechin to prevent oxidative processes of proteins. The carbonylation of each particular protein was quantified by combining the labelling of protein carbonyls by fluorescein-5-thiosemicarbazide (FTSC) with 1-D or 2-D gel electrophoresis. Alpha skeletal actin, glycogen phosphorylase, unnamed protein product (UNP) similar to enolase, pyruvate kinase, isoforms of creatine kinase, aldolase A and an isoform of glyceraldehyde 3-phosphate dehydrogenase (G3PDH) showed elevated oxidation in chilled non-supplemented mince. Myosin heavy chain (MHC) was not carbonylated in chilled muscle, but an extensive MHC degradation was observed in those samples. The supplementation of catechin reduced protein oxidation and lipid oxidation in a concentration-dependent manner: control>25>100≈200ppm. Therefore, the highest catechin concentrations (100 and 200ppm) exhibited the strongest antioxidant activity. Catechin (200ppm) reduced significantly carbonylation of protein spots identified as glycogen phosphorylase, pyruvate kinase muscle isozyme, isoforms of creatine kinase. Conversely, catechin was ineffective to inhibit the oxidation of actin and UNP similar to enolase. These results draw attention to the inefficiency of catechin to prevent actin oxidation, in contrast to the extremely high efficiency of catechin in inhibiting oxidation of lipids and other proteins.
authors.affiliation: Instituto de Investigaciones Marinas, Eduardo Cabello 6, Vigo, Spain. mpazos@iim.csic.es
authors.forename: Manuel
authors.initials: M
authors.lastname: Pazos
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Maestre
authors.forename: José M
authors.initials: JM
authors.lastname: Gallardo
authors.forename: Isabel
authors.initials: I
authors.lastname: Medina
mesh_terms: D000818:Animals; D002392:Catechin / Q000494:pharmacology*; D005396:Fish Products / Q000032:analysis*; D029941:Fish Proteins / Q000737:chemistry*; D005511:Food Handling; D010084:Oxidation-Reduction; D010473:Perciformes; D050050:Protein Carbonylation; D040901:Proteomics; D017747:Seafood / Q000032:analysis*
chemical_list: D029941:Fish Proteins; D002392:Catechin