Fish proteins as targets of ferrous-catalyzed oxidation: Identification of protein carbonyls by fluorescent labeling on two-dimensional gels and MALDI-TOF/TOF mass spectrometry
15 pages, 4 figures Protein oxidation in fish meat is considered to affect negatively the muscle texture. An important source of free radicals taking part in this process is Fenton’s reaction dependent on ferrous ions present in the tissue. The aim of this study was to investigate the susceptibility...
Published in: | Journal of Agricultural and Food Chemistry |
---|---|
Main Authors: | , , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
American Chemical Society
2011
|
Subjects: | |
Online Access: | http://hdl.handle.net/10261/279861 https://doi.org/10.1021/jf201080t |
id |
ftcsic:oai:digital.csic.es:10261/279861 |
---|---|
record_format |
openpolar |
spelling |
ftcsic:oai:digital.csic.es:10261/279861 2024-02-11T10:03:57+01:00 Fish proteins as targets of ferrous-catalyzed oxidation: Identification of protein carbonyls by fluorescent labeling on two-dimensional gels and MALDI-TOF/TOF mass spectrometry Pazos, Manuel Pereira da Rocha, Ángela Roepstorff, Peter Rogowska-Wrzesinska, Adelina 2011 http://hdl.handle.net/10261/279861 https://doi.org/10.1021/jf201080t en eng American Chemical Society https://doi.org/10.1021/jf201080t Sí Journal of Agricultural and Food Chemistry 59(14): 7962–7977 (2011) 0021-8561 http://hdl.handle.net/10261/279861 doi:10.1021/jf201080t 1520-5118 none Protein oxidation Protein carbonyls Metal-catalyzed protein oxidation Gadus morhua Fluorescein-5-thiosemicarbazide (FTSC) MALDI-TOF/TOF Proteomics artículo 2011 ftcsic https://doi.org/10.1021/jf201080t 2024-01-16T11:29:21Z 15 pages, 4 figures Protein oxidation in fish meat is considered to affect negatively the muscle texture. An important source of free radicals taking part in this process is Fenton’s reaction dependent on ferrous ions present in the tissue. The aim of this study was to investigate the susceptibility of cod muscle proteins in sarcoplasmic and myofibril fractions to in vitro metal-catalyzed oxidation and to point out protein candidates that might play a major role in the deterioration of fish quality. Extracted control proteins and proteins subjected to free radicals generated by Fe(II)/ascorbate mixture were labeled with fluorescein-5-thiosemicarbazide (FTSC) to tag carbonyl groups and separated by two-dimensional gel electrophoresis. Consecutive visualization of protein carbonyl levels by capturing the FTSC signal and total protein levels by capturing the SyproRuby staining signal allowed us to quantify the relative change in protein carbonyl levels corrected for changes in protein content. Proteins were identified using MALDI-TOF/TOF mass spectrometry and homology-based searches. The results show that freshly extracted cod muscle proteins exhibit a detectable carbonylation background and that the incubation with Fe(II)/ascorbate triggers a further oxidation of both sarcoplasmic and myofibril proteins. Different proteins exhibited various degrees of sensitivity to oxidation processes. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), nucleoside diphosphate kinase B (NDK), triosephosphate isomerase, phosphoglycerate mutase, lactate dehydrogenase, creatine kinase, and enolase were the sarcoplasmic proteins most vulnerable to ferrous-catalyzed oxidation. Moreover, NDK, phosphoglycerate mutase, and GAPDH were identified in several spots differing by their pI, and those forms showed different susceptibilities to metal-catalyzed oxidation, indicating that post-translational modifications may change the resistance of proteins to oxidative damage. The Fe(II)/ascorbate treatment significantly increased carbonylation of ... Article in Journal/Newspaper Gadus morhua Digital.CSIC (Spanish National Research Council) Journal of Agricultural and Food Chemistry 59 14 7962 7977 |
institution |
Open Polar |
collection |
Digital.CSIC (Spanish National Research Council) |
op_collection_id |
ftcsic |
language |
English |
topic |
Protein oxidation Protein carbonyls Metal-catalyzed protein oxidation Gadus morhua Fluorescein-5-thiosemicarbazide (FTSC) MALDI-TOF/TOF Proteomics |
spellingShingle |
Protein oxidation Protein carbonyls Metal-catalyzed protein oxidation Gadus morhua Fluorescein-5-thiosemicarbazide (FTSC) MALDI-TOF/TOF Proteomics Pazos, Manuel Pereira da Rocha, Ángela Roepstorff, Peter Rogowska-Wrzesinska, Adelina Fish proteins as targets of ferrous-catalyzed oxidation: Identification of protein carbonyls by fluorescent labeling on two-dimensional gels and MALDI-TOF/TOF mass spectrometry |
topic_facet |
Protein oxidation Protein carbonyls Metal-catalyzed protein oxidation Gadus morhua Fluorescein-5-thiosemicarbazide (FTSC) MALDI-TOF/TOF Proteomics |
description |
15 pages, 4 figures Protein oxidation in fish meat is considered to affect negatively the muscle texture. An important source of free radicals taking part in this process is Fenton’s reaction dependent on ferrous ions present in the tissue. The aim of this study was to investigate the susceptibility of cod muscle proteins in sarcoplasmic and myofibril fractions to in vitro metal-catalyzed oxidation and to point out protein candidates that might play a major role in the deterioration of fish quality. Extracted control proteins and proteins subjected to free radicals generated by Fe(II)/ascorbate mixture were labeled with fluorescein-5-thiosemicarbazide (FTSC) to tag carbonyl groups and separated by two-dimensional gel electrophoresis. Consecutive visualization of protein carbonyl levels by capturing the FTSC signal and total protein levels by capturing the SyproRuby staining signal allowed us to quantify the relative change in protein carbonyl levels corrected for changes in protein content. Proteins were identified using MALDI-TOF/TOF mass spectrometry and homology-based searches. The results show that freshly extracted cod muscle proteins exhibit a detectable carbonylation background and that the incubation with Fe(II)/ascorbate triggers a further oxidation of both sarcoplasmic and myofibril proteins. Different proteins exhibited various degrees of sensitivity to oxidation processes. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), nucleoside diphosphate kinase B (NDK), triosephosphate isomerase, phosphoglycerate mutase, lactate dehydrogenase, creatine kinase, and enolase were the sarcoplasmic proteins most vulnerable to ferrous-catalyzed oxidation. Moreover, NDK, phosphoglycerate mutase, and GAPDH were identified in several spots differing by their pI, and those forms showed different susceptibilities to metal-catalyzed oxidation, indicating that post-translational modifications may change the resistance of proteins to oxidative damage. The Fe(II)/ascorbate treatment significantly increased carbonylation of ... |
format |
Article in Journal/Newspaper |
author |
Pazos, Manuel Pereira da Rocha, Ángela Roepstorff, Peter Rogowska-Wrzesinska, Adelina |
author_facet |
Pazos, Manuel Pereira da Rocha, Ángela Roepstorff, Peter Rogowska-Wrzesinska, Adelina |
author_sort |
Pazos, Manuel |
title |
Fish proteins as targets of ferrous-catalyzed oxidation: Identification of protein carbonyls by fluorescent labeling on two-dimensional gels and MALDI-TOF/TOF mass spectrometry |
title_short |
Fish proteins as targets of ferrous-catalyzed oxidation: Identification of protein carbonyls by fluorescent labeling on two-dimensional gels and MALDI-TOF/TOF mass spectrometry |
title_full |
Fish proteins as targets of ferrous-catalyzed oxidation: Identification of protein carbonyls by fluorescent labeling on two-dimensional gels and MALDI-TOF/TOF mass spectrometry |
title_fullStr |
Fish proteins as targets of ferrous-catalyzed oxidation: Identification of protein carbonyls by fluorescent labeling on two-dimensional gels and MALDI-TOF/TOF mass spectrometry |
title_full_unstemmed |
Fish proteins as targets of ferrous-catalyzed oxidation: Identification of protein carbonyls by fluorescent labeling on two-dimensional gels and MALDI-TOF/TOF mass spectrometry |
title_sort |
fish proteins as targets of ferrous-catalyzed oxidation: identification of protein carbonyls by fluorescent labeling on two-dimensional gels and maldi-tof/tof mass spectrometry |
publisher |
American Chemical Society |
publishDate |
2011 |
url |
http://hdl.handle.net/10261/279861 https://doi.org/10.1021/jf201080t |
genre |
Gadus morhua |
genre_facet |
Gadus morhua |
op_relation |
https://doi.org/10.1021/jf201080t Sí Journal of Agricultural and Food Chemistry 59(14): 7962–7977 (2011) 0021-8561 http://hdl.handle.net/10261/279861 doi:10.1021/jf201080t 1520-5118 |
op_rights |
none |
op_doi |
https://doi.org/10.1021/jf201080t |
container_title |
Journal of Agricultural and Food Chemistry |
container_volume |
59 |
container_issue |
14 |
container_start_page |
7962 |
op_container_end_page |
7977 |
_version_ |
1790600356144611328 |