Investigation by Synchrotron Radiation Circular Dichroism of the Secondary Structure Evolution of Pepsin under Oxidative Environment
Food processing affects the structure and chemical state of proteins. In particular, protein oxidation occurs and may impair protein properties. These chemical reactions initiated during processing can develop during digestion. Indeed, the physicochemical conditions of the stomach (oxygen pressure,...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-05-01
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| Series: | Foods |
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| Online Access: | https://www.mdpi.com/2304-8158/10/5/998 |
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| author | Laetitia Théron Aline Bonifacie Jérémy Delabre Thierry Sayd Laurent Aubry Philippe Gatellier Christine Ravel Christophe Chambon Thierry Astruc Jacques Rouel Véronique Santé-Lhoutellier Matthieu Réfrégiers Frank Wien |
| author_facet | Laetitia Théron Aline Bonifacie Jérémy Delabre Thierry Sayd Laurent Aubry Philippe Gatellier Christine Ravel Christophe Chambon Thierry Astruc Jacques Rouel Véronique Santé-Lhoutellier Matthieu Réfrégiers Frank Wien |
| author_sort | Laetitia Théron |
| collection | DOAJ |
| description | Food processing affects the structure and chemical state of proteins. In particular, protein oxidation occurs and may impair protein properties. These chemical reactions initiated during processing can develop during digestion. Indeed, the physicochemical conditions of the stomach (oxygen pressure, low pH) favor oxidation. In that respect, digestive proteases may be affected as well. Yet, very little is known about the link between endogenous oxidation of digestive enzymes, their potential denaturation, and, therefore, food protein digestibility. Thus, the objective of this study is to understand how oxidative chemical processes will impact the pepsin secondary structure and its hydrolytic activity. The folding and unfolding kinetics of pepsin under oxidative conditions was determined using Synchrotron Radiation Circular Dichroism. SRCD gave us the possibility to monitor the rapid kinetics of protein folding and unfolding in real-time, giving highly resolved spectral data. The proteolytic activity of control and oxidized pepsin was investigated by MALDI-TOF mass spectrometry on a meat protein model, the creatine kinase. MALDI-TOF MS allowed a rapid evaluation of the proteolytic activity through peptide fingerprint. This study opens up new perspectives by shifting the digestion paradigm taking into account the gastric digestive enzyme and its substrate. |
| first_indexed | 2024-03-10T11:44:41Z |
| format | Article |
| id | doaj.art-d84e111d53a1410c844f077997d5dfcb |
| institution | Directory Open Access Journal |
| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-03-10T11:44:41Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Foods |
| spelling | doaj.art-d84e111d53a1410c844f077997d5dfcb2023-11-21T18:11:45ZengMDPI AGFoods2304-81582021-05-0110599810.3390/foods10050998Investigation by Synchrotron Radiation Circular Dichroism of the Secondary Structure Evolution of Pepsin under Oxidative EnvironmentLaetitia Théron0Aline Bonifacie1Jérémy Delabre2Thierry Sayd3Laurent Aubry4Philippe Gatellier5Christine Ravel6Christophe Chambon7Thierry Astruc8Jacques Rouel9Véronique Santé-Lhoutellier10Matthieu Réfrégiers11Frank Wien12Institut National de Recherche Pour L’agriculture, L’alimentation et L’environnement (INRAE), UR370 Qualité des Produits Animaux, F-63122 Saint Genès-Champanelle, FranceInstitut National de Recherche Pour L’agriculture, L’alimentation et L’environnement (INRAE), UR370 Qualité des Produits Animaux, F-63122 Saint Genès-Champanelle, FranceInstitut National de Recherche Pour L’agriculture, L’alimentation et L’environnement (INRAE), Plateforme D’exploration du Métabolisme Composante Protéomique (PFEMcp), F-63122 Saint Genès-Champanelle, FranceInstitut National de Recherche Pour L’agriculture, L’alimentation et L’environnement (INRAE), UR370 Qualité des Produits Animaux, F-63122 Saint Genès-Champanelle, FranceInstitut National de Recherche Pour L’agriculture, L’alimentation et L’environnement (INRAE), UR370 Qualité des Produits Animaux, F-63122 Saint Genès-Champanelle, FranceInstitut National de Recherche Pour L’agriculture, L’alimentation et L’environnement (INRAE), UR370 Qualité des Produits Animaux, F-63122 Saint Genès-Champanelle, FranceInstitut National de Recherche Pour L’agriculture, L’alimentation et L’environnement (INRAE), UR370 Qualité des Produits Animaux, F-63122 Saint Genès-Champanelle, FranceInstitut National de Recherche Pour L’agriculture, L’alimentation et L’environnement (INRAE), Plateforme D’exploration du Métabolisme Composante Protéomique (PFEMcp), F-63122 Saint Genès-Champanelle, FranceInstitut National de Recherche Pour L’agriculture, L’alimentation et L’environnement (INRAE), UR370 Qualité des Produits Animaux, F-63122 Saint Genès-Champanelle, FranceInstitut National de Recherche Pour L’agriculture, L’alimentation et L’environnement (INRAE), UR370 Qualité des Produits Animaux, F-63122 Saint Genès-Champanelle, FranceInstitut National de Recherche Pour L’agriculture, L’alimentation et L’environnement (INRAE), UR370 Qualité des Produits Animaux, F-63122 Saint Genès-Champanelle, FranceDISCO Beamline, Synchrotron SOLEIL, F-91192 Gif-sur-Yvette, FranceDISCO Beamline, Synchrotron SOLEIL, F-91192 Gif-sur-Yvette, FranceFood processing affects the structure and chemical state of proteins. In particular, protein oxidation occurs and may impair protein properties. These chemical reactions initiated during processing can develop during digestion. Indeed, the physicochemical conditions of the stomach (oxygen pressure, low pH) favor oxidation. In that respect, digestive proteases may be affected as well. Yet, very little is known about the link between endogenous oxidation of digestive enzymes, their potential denaturation, and, therefore, food protein digestibility. Thus, the objective of this study is to understand how oxidative chemical processes will impact the pepsin secondary structure and its hydrolytic activity. The folding and unfolding kinetics of pepsin under oxidative conditions was determined using Synchrotron Radiation Circular Dichroism. SRCD gave us the possibility to monitor the rapid kinetics of protein folding and unfolding in real-time, giving highly resolved spectral data. The proteolytic activity of control and oxidized pepsin was investigated by MALDI-TOF mass spectrometry on a meat protein model, the creatine kinase. MALDI-TOF MS allowed a rapid evaluation of the proteolytic activity through peptide fingerprint. This study opens up new perspectives by shifting the digestion paradigm taking into account the gastric digestive enzyme and its substrate.https://www.mdpi.com/2304-8158/10/5/998protein oxidationsynchrotron radiation circular dichroismdigestionpepsinmass spectrometry |
| spellingShingle | Laetitia Théron Aline Bonifacie Jérémy Delabre Thierry Sayd Laurent Aubry Philippe Gatellier Christine Ravel Christophe Chambon Thierry Astruc Jacques Rouel Véronique Santé-Lhoutellier Matthieu Réfrégiers Frank Wien Investigation by Synchrotron Radiation Circular Dichroism of the Secondary Structure Evolution of Pepsin under Oxidative Environment Foods protein oxidation synchrotron radiation circular dichroism digestion pepsin mass spectrometry |
| title | Investigation by Synchrotron Radiation Circular Dichroism of the Secondary Structure Evolution of Pepsin under Oxidative Environment |
| title_full | Investigation by Synchrotron Radiation Circular Dichroism of the Secondary Structure Evolution of Pepsin under Oxidative Environment |
| title_fullStr | Investigation by Synchrotron Radiation Circular Dichroism of the Secondary Structure Evolution of Pepsin under Oxidative Environment |
| title_full_unstemmed | Investigation by Synchrotron Radiation Circular Dichroism of the Secondary Structure Evolution of Pepsin under Oxidative Environment |
| title_short | Investigation by Synchrotron Radiation Circular Dichroism of the Secondary Structure Evolution of Pepsin under Oxidative Environment |
| title_sort | investigation by synchrotron radiation circular dichroism of the secondary structure evolution of pepsin under oxidative environment |
| topic | protein oxidation synchrotron radiation circular dichroism digestion pepsin mass spectrometry |
| url | https://www.mdpi.com/2304-8158/10/5/998 |
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