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Papers by Michel Seve
pre-analytical step for complex eukaryotic samples fractionation compatible with quantitative iTR... more pre-analytical step for complex eukaryotic samples fractionation compatible with quantitative iTRAQ labeling.
Annales De Biologie Clinique, 2007
Cet article de synthese se penche sur une nouvelle discipline biologique et medicale, la « proteo... more Cet article de synthese se penche sur une nouvelle discipline biologique et medicale, la « proteomique clinique ». Cette approche « post-genomique » vise a utiliser l’etude du proteome, c’est-a-dire de l’ensemble des peptides et proteines presents dans un echantillon biologique, pour donner une information diagnostique, pronostique ou de suivi therapeutique des pathologies humaines. Cette nouvelle discipline est en plein developpement car elle presente d’importantes perspectives pour les pathologies complexes ou pour la detection precoce de pathologies telles que les cancers. Les elements de cette revue resultent du travail d’un groupe thematique de la SFBC (Societe francaise de biologie clinique) de 2004 a 2006 dont l’objectif etait d’evaluer l’etat actuel de la « proteomique clinique » et de s’interroger sur son developpement et son application future pour les biologistes. Il s’agit donc d’un etat des lieux assez large decrivant les approches deja accessibles dans les laboratoires...
Archives of Cardiovascular Diseases Supplements, 2021
Introduction The secretome is a set of proteins secreted by a cell at a given time and under cert... more Introduction The secretome is a set of proteins secreted by a cell at a given time and under certain conditions. The secretome, easily accessible from plasma and analyzable by a proteomic approach, represents a good opportunity to identify biomarkers in septic shock. This pathology corresponds to a systemic inflammatory response syndrome related to an infection and associated with cardiocirculatory failure leading to organ dysfunction with a mortality rate of 40%. The identification of diagnostic/prognostic biomarkers is crucial to improve patient outcome. Objective Study the secretome of a rat model of endotoxemic shock to identify relevant deregulated proteins (DEP). Methods Endotoxemic shock was induced by lipopolysaccharides injection (LPS, S. enterica typhi, 0.5 mg/kg, iv) and compared to controls (Ringer lactate, iv). Under isoflurane anesthesia, carotid cannulation allowed evaluation of mean arterial blood pressure (MAP), heart rate and blood sampling (T0, and T50 or T90 min ...
International Journal of Molecular Sciences, 2021
Mutations in the X-linked MECP2 gene are responsible for Rett syndrome (RTT), a severe neurologic... more Mutations in the X-linked MECP2 gene are responsible for Rett syndrome (RTT), a severe neurological disorder. MECP2 is a transcriptional modulator that finely regulates the expression of many genes, specifically in the central nervous system. Several studies have functionally linked the loss of MECP2 in astrocytes to the appearance and progression of the RTT phenotype in a non-cell autonomous manner and mechanisms are still unknown. Here, we used primary astroglial cells from Mecp2-deficient (KO) pups to identify deregulated secreted proteins. Using a differential quantitative proteomic analysis, twenty-nine proteins have been identified and four were confirmed by Western blotting with new samples as significantly deregulated. To further verify the functional relevance of these proteins in RTT, we tested their effects on the dendritic morphology of primary cortical neurons from Mecp2 KO mice that are known to display shorter dendritic processes. Using Sholl analysis, we found that i...
Journal of Proteomics, 2019
title Click here to download Manuscript: 05-R_AcGef_01_Title-page-revision.docx Click here to vie... more title Click here to download Manuscript: 05-R_AcGef_01_Title-page-revision.docx Click here to view linked References 42 44 57 proteins: upstream neighbors of apoptosis Metabolic process Cell cycle/Chromosome segregation Development process /Cell death … proteins
Asian Pacific journal of allergy and immunology, Jan 15, 2018
Tree pollens are well-known aeroallergens all over the world. Little is known about the allergeni... more Tree pollens are well-known aeroallergens all over the world. Little is known about the allergenicity of Morus alba (white mulberry) pollen. We aimed to explore the potential allergens of this pollen and its clinical relevance in tree pollen allergic patients living in Istanbul, Turkey. Twenty three seasonal allergic rhinitis patients with a confirmed tree pollen allergy and 5 healthy control subjects underwent skin prick and nasal provocation tests with M.alba pollen extract. The pollen extract was then resolved by gel electrophoresis, and immunoblotted with sera from patients/control individuals to detect the potential allergenic proteins. The prevalent IgE binding proteins from 1D-gel were analyzed by MALDI-TOF/TOF. Eleven out of 23 patients were reactive to the extract with skin prick tests. Seven of those patients also reacted positively to the nasal provocation tests. The most common IgE-binding pollen proteins were detected between 55-100 kDa, and also at molecular weights lo...
Expert Review of Proteomics, 2018
Introduction: Acetylation is a widely occurring post-translational modification (PTM) of proteins... more Introduction: Acetylation is a widely occurring post-translational modification (PTM) of proteins that plays a crucial role in many cellular physiological and pathological processes. Over the last decade, acetylation analyses required the development of multiple methods to target individual acetylated proteins, as well as to cover a broader description of acetylated proteins that comprise the acetylome. Areas covered: This review discusses the different types of acetylation (N-ter/K-/O-acetylation) and then describes some major strategies that have been reported in the literature to detect, enrich, identify and quantify protein acetylation. The review highlights the advantages and limitations of these strategies, to guide researchers in designing their experimental investigations and analysis of protein acetylation. Finally, this review highlights the main applications of acetylomics (proteomics based on mass spectrometry) for understanding physiological and pathological conditions. Expert opinion: Recent advances in acetylomics have enhanced knowledge of the biological and pathological roles of protein acetylation and the acetylome. Besides, radiolabeling and western blotting remain also techniques-of-choice for targeted protein acetylation. Future challenges in acetylomics to analyze the N-ter and K-acetylome will most likely require enrichment/fractionation, MS instrumentation and bioinformatics. Challenges also remain to identify the potential biological roles of O-acetylation and cross-talk with other PTMs.
Journal of biochemical and molecular toxicology, 2018
Investigating new antimicrobial and antiparasitic components from Viperidae venoms represents an ... more Investigating new antimicrobial and antiparasitic components from Viperidae venoms represents an alternative therapeutic strategy. In this study, we report the characterization of a disintegrin isolated from Cerastes cerastes venom, exhibiting antiparasitic activity on Leishmania infantum promastigotes. Indeed, isolated disintegrin, referred to Disintegrin_Cc, induced 84.75% of parasiticidal activity and deep morphological alterations on the parasites. SDS-PAGE analysis indicated that this disintegrin was homogenous. This dimeric disintegrin of 14,193.97 Da contains an RGD domain and four intramolecular disulfide bridges. It presents a high percentage of identity with other related snake disintegrins. Predicted 3D structure indicated that this peptide shares partial homology with well-known active antimicrobial peptides. Disintegrin_Cc inhibited 80% of arachidonic acid-induced platelet aggregation. The obtained results suggest that the isolated molecule plays a dual role as a disint...
Environmental science and pollution research international, 2018
Over the last decades, engineered nanomaterials have been widely used in various applications due... more Over the last decades, engineered nanomaterials have been widely used in various applications due to their interesting properties. Among them, iron oxide nanoparticles (IONPs) are used as theranostic agents for cancer, and also as contrast agents in magnetic resonance imaging. With the increasing production and use of these IONPs, there is an evident raise of IONP exposure and subsequently a higher risk of adverse outcome for humans and the environment. In this work, we aimed to investigate the effects of sub-acute IONP exposure on Wistar rat, particularly (i) on the emotional and learning/memory behavior, (ii) on the hematological and biochemical parameters, (iii) on the neurotransmitter content, and (vi) on the trace element homeostasis. Rats were treated during seven consecutive days by intranasal instillations at a dose of 10 mg/kg body weight. The mean body weight increased significantly in IONP-exposed rats. Moreover, several hematological parameters were normal in treated rat...
Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements (GMS), 2018
Engineered nanomaterials are used in various applications due to their particular properties. Amo... more Engineered nanomaterials are used in various applications due to their particular properties. Among them, Iron Oxide Nanoparticles (FeO-NPs) are used in Biomedicine as theranostic agents i.e. contrast agents in Magnetic Resonance Imaging and cancer treatment. With the increasing production and use of these FeO-NPs, there is an evident raise of FeO-NPs exposure and subsequently a higher risk of adverse outcomes for the environment and Human. In the present paper, we investigated the effects of an intravenous daily FeO-NPs exposure on Wistar rat for one week. As results, we showed that several hematological parameters and transaminase (ALT and AST) levels as well as organ histology remained unchanged in treated rats. Neither the catecholamine levels nor the emotional behavior and learning / memory capacities of rats were impacted by the sub-acute intravenous exposure to FeO-NPs. However, iron level in plasma and iron content homeostasis in brain were disrupted after this exposure. Thu...
The journal of venomous animals and toxins including tropical diseases, 2018
Sperm contains a wealth of cell surface receptors and ion channels that are required for most of ... more Sperm contains a wealth of cell surface receptors and ion channels that are required for most of its basic functions such as motility and acrosome reaction. Conversely, animal venoms are enriched in bioactive compounds that primarily target those ion channels and cell surface receptors. We hypothesized, therefore, that animal venoms should be rich enough in sperm-modulating compounds for a drug discovery program. Our objective was to demonstrate this fact by using a sperm-based phenotypic screening to identify positive modulators from the venom of . Herein, as proof of concept that venoms contain interesting compounds for sperm physiology, we fractionated snake venom by RP-HPLC and screened for bioactive fractions capable of accelerating mouse sperm motility (primary screening). Next, we purified each compound from the positive fraction by cation exchange and identified the bioactive peptide by secondary screening. The peptide sequence was established by Edman sequencing of the redu...
Frontiers in immunology, 2017
Bronchiolitis obliterans syndrome (BOS), the main manifestation of chronic lung allograft dysfunc... more Bronchiolitis obliterans syndrome (BOS), the main manifestation of chronic lung allograft dysfunction, leads to poor long-term survival after lung transplantation. Identifying predictors of BOS is essential to prevent the progression of dysfunction before irreversible damage occurs. By using a large set of 107 samples from lung recipients, we performed microarray gene expression profiling of whole blood to identify early biomarkers of BOS, including samples from 49 patients with stable function for at least 3 years, 32 samples collected at least 6 months before BOS diagnosis (prediction group), and 26 samples at or after BOS diagnosis (diagnosis group). An independent set from 25 lung recipients was used for validation by quantitative PCR (13 stables, 11 in the prediction group, and 8 in the diagnosis group). We identified 50 transcripts differentially expressed between stable and BOS recipients. Three genes, namely POU class 2 associating factor 1 (POU2AF1), T-cell leukemia/lymphom...
Clinical Science, 2017
Background: Exercise and citrulline (CIT) are both regulators of muscle protein metabolism. Howev... more Background: Exercise and citrulline (CIT) are both regulators of muscle protein metabolism. However, the combination of both has been under-studied yet may have synergistic effects on muscle metabolism and performance. Methods: Three-month-old healthy male rats were randomly assigned to be fed ad libitum for 4 weeks with either a citrulline-enriched diet (1 g·kg−1·day−1) (CIT) or an isonitrogenous standard diet (by addition of nonessential amino acid) (Ctrl) and trained (running on treadmill 5 days·week−1) (ex) or not. Maximal endurance activity and body composition were assessed, and muscle protein metabolism (protein synthesis, proteomic approach) and energy metabolism [energy expenditure, mitochondrial metabolism] were explored. Results: Body composition was affected by exercise but not by CIT supplementation. Endurance training was associated with a higher maximal endurance capacity than sedentary groups (P<0.001), and running time was 14% higher in the CITex group than the C...
ELECTROPHORESIS, 2017
High-throughput mass spectrometry-based proteomic analysis requires peptide fractionation to simp... more High-throughput mass spectrometry-based proteomic analysis requires peptide fractionation to simplify complex biological samples and increase proteome coverage. OFFGEL fractionation technology became a common method to separate peptides or proteins using isoelectric focusing in an immobilized pH gradient. However, the OFFGEL focusing process may be further optimized and controlled in terms of separation time and pI resolution. Here, we evaluated OFFGEL technology to separate peptides from different samples in the presence of lowmolecular-weight (LMW) color pI markers to visualize the focusing process. LMW color pI markers covering a large pH range were added to the peptide mixture before OFFGEL fractionation using a 24-wells device encompassing the pH range 3-10. We also explored the impact of LMW color pI markers on peptide fractionation labeled previously for iTRAQ. Then, fractionated peptides were separated by RP_HPLC prior to MS analysis using MALDI-TOF/TOF mass spectrometry in MS and MS/MS modes. Here we report the performance of the peptide focusing process in the presence of LMW color pI markers as on-line trackers during the OFFGEL process and the possibility to use them as pI controls for peptide focusing. This method improves the workflow for peptide fractionation in a bottom-up proteomic approach with or without iTRAQ labeling.
PROTEOMICS, 2016
Besides their main contribution as substrates for protein synthesis, amino acids as signaling mol... more Besides their main contribution as substrates for protein synthesis, amino acids as signaling molecules could exert some regulatory functions on protein synthesis and/or proteolysis that have been emphasized in a number of recent studies. Several publications have highlighted supplemental roles of those amino acids in protein metabolism as well as in immunity, heat shock response, or apoptosis processes. In this way, via their regulatory properties, selected amino acids (such as leucine, glutamine, arginine, citrulline, or methionine) directly influence the proteome. In this review, we are proposing an overview of the regulation of the proteome by amino acids in mammals.
pre-analytical step for complex eukaryotic samples fractionation compatible with quantitative iTR... more pre-analytical step for complex eukaryotic samples fractionation compatible with quantitative iTRAQ labeling.
Annales De Biologie Clinique, 2007
Cet article de synthese se penche sur une nouvelle discipline biologique et medicale, la « proteo... more Cet article de synthese se penche sur une nouvelle discipline biologique et medicale, la « proteomique clinique ». Cette approche « post-genomique » vise a utiliser l’etude du proteome, c’est-a-dire de l’ensemble des peptides et proteines presents dans un echantillon biologique, pour donner une information diagnostique, pronostique ou de suivi therapeutique des pathologies humaines. Cette nouvelle discipline est en plein developpement car elle presente d’importantes perspectives pour les pathologies complexes ou pour la detection precoce de pathologies telles que les cancers. Les elements de cette revue resultent du travail d’un groupe thematique de la SFBC (Societe francaise de biologie clinique) de 2004 a 2006 dont l’objectif etait d’evaluer l’etat actuel de la « proteomique clinique » et de s’interroger sur son developpement et son application future pour les biologistes. Il s’agit donc d’un etat des lieux assez large decrivant les approches deja accessibles dans les laboratoires...
Archives of Cardiovascular Diseases Supplements, 2021
Introduction The secretome is a set of proteins secreted by a cell at a given time and under cert... more Introduction The secretome is a set of proteins secreted by a cell at a given time and under certain conditions. The secretome, easily accessible from plasma and analyzable by a proteomic approach, represents a good opportunity to identify biomarkers in septic shock. This pathology corresponds to a systemic inflammatory response syndrome related to an infection and associated with cardiocirculatory failure leading to organ dysfunction with a mortality rate of 40%. The identification of diagnostic/prognostic biomarkers is crucial to improve patient outcome. Objective Study the secretome of a rat model of endotoxemic shock to identify relevant deregulated proteins (DEP). Methods Endotoxemic shock was induced by lipopolysaccharides injection (LPS, S. enterica typhi, 0.5 mg/kg, iv) and compared to controls (Ringer lactate, iv). Under isoflurane anesthesia, carotid cannulation allowed evaluation of mean arterial blood pressure (MAP), heart rate and blood sampling (T0, and T50 or T90 min ...
International Journal of Molecular Sciences, 2021
Mutations in the X-linked MECP2 gene are responsible for Rett syndrome (RTT), a severe neurologic... more Mutations in the X-linked MECP2 gene are responsible for Rett syndrome (RTT), a severe neurological disorder. MECP2 is a transcriptional modulator that finely regulates the expression of many genes, specifically in the central nervous system. Several studies have functionally linked the loss of MECP2 in astrocytes to the appearance and progression of the RTT phenotype in a non-cell autonomous manner and mechanisms are still unknown. Here, we used primary astroglial cells from Mecp2-deficient (KO) pups to identify deregulated secreted proteins. Using a differential quantitative proteomic analysis, twenty-nine proteins have been identified and four were confirmed by Western blotting with new samples as significantly deregulated. To further verify the functional relevance of these proteins in RTT, we tested their effects on the dendritic morphology of primary cortical neurons from Mecp2 KO mice that are known to display shorter dendritic processes. Using Sholl analysis, we found that i...
Journal of Proteomics, 2019
title Click here to download Manuscript: 05-R_AcGef_01_Title-page-revision.docx Click here to vie... more title Click here to download Manuscript: 05-R_AcGef_01_Title-page-revision.docx Click here to view linked References 42 44 57 proteins: upstream neighbors of apoptosis Metabolic process Cell cycle/Chromosome segregation Development process /Cell death … proteins
Asian Pacific journal of allergy and immunology, Jan 15, 2018
Tree pollens are well-known aeroallergens all over the world. Little is known about the allergeni... more Tree pollens are well-known aeroallergens all over the world. Little is known about the allergenicity of Morus alba (white mulberry) pollen. We aimed to explore the potential allergens of this pollen and its clinical relevance in tree pollen allergic patients living in Istanbul, Turkey. Twenty three seasonal allergic rhinitis patients with a confirmed tree pollen allergy and 5 healthy control subjects underwent skin prick and nasal provocation tests with M.alba pollen extract. The pollen extract was then resolved by gel electrophoresis, and immunoblotted with sera from patients/control individuals to detect the potential allergenic proteins. The prevalent IgE binding proteins from 1D-gel were analyzed by MALDI-TOF/TOF. Eleven out of 23 patients were reactive to the extract with skin prick tests. Seven of those patients also reacted positively to the nasal provocation tests. The most common IgE-binding pollen proteins were detected between 55-100 kDa, and also at molecular weights lo...
Expert Review of Proteomics, 2018
Introduction: Acetylation is a widely occurring post-translational modification (PTM) of proteins... more Introduction: Acetylation is a widely occurring post-translational modification (PTM) of proteins that plays a crucial role in many cellular physiological and pathological processes. Over the last decade, acetylation analyses required the development of multiple methods to target individual acetylated proteins, as well as to cover a broader description of acetylated proteins that comprise the acetylome. Areas covered: This review discusses the different types of acetylation (N-ter/K-/O-acetylation) and then describes some major strategies that have been reported in the literature to detect, enrich, identify and quantify protein acetylation. The review highlights the advantages and limitations of these strategies, to guide researchers in designing their experimental investigations and analysis of protein acetylation. Finally, this review highlights the main applications of acetylomics (proteomics based on mass spectrometry) for understanding physiological and pathological conditions. Expert opinion: Recent advances in acetylomics have enhanced knowledge of the biological and pathological roles of protein acetylation and the acetylome. Besides, radiolabeling and western blotting remain also techniques-of-choice for targeted protein acetylation. Future challenges in acetylomics to analyze the N-ter and K-acetylome will most likely require enrichment/fractionation, MS instrumentation and bioinformatics. Challenges also remain to identify the potential biological roles of O-acetylation and cross-talk with other PTMs.
Journal of biochemical and molecular toxicology, 2018
Investigating new antimicrobial and antiparasitic components from Viperidae venoms represents an ... more Investigating new antimicrobial and antiparasitic components from Viperidae venoms represents an alternative therapeutic strategy. In this study, we report the characterization of a disintegrin isolated from Cerastes cerastes venom, exhibiting antiparasitic activity on Leishmania infantum promastigotes. Indeed, isolated disintegrin, referred to Disintegrin_Cc, induced 84.75% of parasiticidal activity and deep morphological alterations on the parasites. SDS-PAGE analysis indicated that this disintegrin was homogenous. This dimeric disintegrin of 14,193.97 Da contains an RGD domain and four intramolecular disulfide bridges. It presents a high percentage of identity with other related snake disintegrins. Predicted 3D structure indicated that this peptide shares partial homology with well-known active antimicrobial peptides. Disintegrin_Cc inhibited 80% of arachidonic acid-induced platelet aggregation. The obtained results suggest that the isolated molecule plays a dual role as a disint...
Environmental science and pollution research international, 2018
Over the last decades, engineered nanomaterials have been widely used in various applications due... more Over the last decades, engineered nanomaterials have been widely used in various applications due to their interesting properties. Among them, iron oxide nanoparticles (IONPs) are used as theranostic agents for cancer, and also as contrast agents in magnetic resonance imaging. With the increasing production and use of these IONPs, there is an evident raise of IONP exposure and subsequently a higher risk of adverse outcome for humans and the environment. In this work, we aimed to investigate the effects of sub-acute IONP exposure on Wistar rat, particularly (i) on the emotional and learning/memory behavior, (ii) on the hematological and biochemical parameters, (iii) on the neurotransmitter content, and (vi) on the trace element homeostasis. Rats were treated during seven consecutive days by intranasal instillations at a dose of 10 mg/kg body weight. The mean body weight increased significantly in IONP-exposed rats. Moreover, several hematological parameters were normal in treated rat...
Journal of trace elements in medicine and biology : organ of the Society for Minerals and Trace Elements (GMS), 2018
Engineered nanomaterials are used in various applications due to their particular properties. Amo... more Engineered nanomaterials are used in various applications due to their particular properties. Among them, Iron Oxide Nanoparticles (FeO-NPs) are used in Biomedicine as theranostic agents i.e. contrast agents in Magnetic Resonance Imaging and cancer treatment. With the increasing production and use of these FeO-NPs, there is an evident raise of FeO-NPs exposure and subsequently a higher risk of adverse outcomes for the environment and Human. In the present paper, we investigated the effects of an intravenous daily FeO-NPs exposure on Wistar rat for one week. As results, we showed that several hematological parameters and transaminase (ALT and AST) levels as well as organ histology remained unchanged in treated rats. Neither the catecholamine levels nor the emotional behavior and learning / memory capacities of rats were impacted by the sub-acute intravenous exposure to FeO-NPs. However, iron level in plasma and iron content homeostasis in brain were disrupted after this exposure. Thu...
The journal of venomous animals and toxins including tropical diseases, 2018
Sperm contains a wealth of cell surface receptors and ion channels that are required for most of ... more Sperm contains a wealth of cell surface receptors and ion channels that are required for most of its basic functions such as motility and acrosome reaction. Conversely, animal venoms are enriched in bioactive compounds that primarily target those ion channels and cell surface receptors. We hypothesized, therefore, that animal venoms should be rich enough in sperm-modulating compounds for a drug discovery program. Our objective was to demonstrate this fact by using a sperm-based phenotypic screening to identify positive modulators from the venom of . Herein, as proof of concept that venoms contain interesting compounds for sperm physiology, we fractionated snake venom by RP-HPLC and screened for bioactive fractions capable of accelerating mouse sperm motility (primary screening). Next, we purified each compound from the positive fraction by cation exchange and identified the bioactive peptide by secondary screening. The peptide sequence was established by Edman sequencing of the redu...
Frontiers in immunology, 2017
Bronchiolitis obliterans syndrome (BOS), the main manifestation of chronic lung allograft dysfunc... more Bronchiolitis obliterans syndrome (BOS), the main manifestation of chronic lung allograft dysfunction, leads to poor long-term survival after lung transplantation. Identifying predictors of BOS is essential to prevent the progression of dysfunction before irreversible damage occurs. By using a large set of 107 samples from lung recipients, we performed microarray gene expression profiling of whole blood to identify early biomarkers of BOS, including samples from 49 patients with stable function for at least 3 years, 32 samples collected at least 6 months before BOS diagnosis (prediction group), and 26 samples at or after BOS diagnosis (diagnosis group). An independent set from 25 lung recipients was used for validation by quantitative PCR (13 stables, 11 in the prediction group, and 8 in the diagnosis group). We identified 50 transcripts differentially expressed between stable and BOS recipients. Three genes, namely POU class 2 associating factor 1 (POU2AF1), T-cell leukemia/lymphom...
Clinical Science, 2017
Background: Exercise and citrulline (CIT) are both regulators of muscle protein metabolism. Howev... more Background: Exercise and citrulline (CIT) are both regulators of muscle protein metabolism. However, the combination of both has been under-studied yet may have synergistic effects on muscle metabolism and performance. Methods: Three-month-old healthy male rats were randomly assigned to be fed ad libitum for 4 weeks with either a citrulline-enriched diet (1 g·kg−1·day−1) (CIT) or an isonitrogenous standard diet (by addition of nonessential amino acid) (Ctrl) and trained (running on treadmill 5 days·week−1) (ex) or not. Maximal endurance activity and body composition were assessed, and muscle protein metabolism (protein synthesis, proteomic approach) and energy metabolism [energy expenditure, mitochondrial metabolism] were explored. Results: Body composition was affected by exercise but not by CIT supplementation. Endurance training was associated with a higher maximal endurance capacity than sedentary groups (P<0.001), and running time was 14% higher in the CITex group than the C...
ELECTROPHORESIS, 2017
High-throughput mass spectrometry-based proteomic analysis requires peptide fractionation to simp... more High-throughput mass spectrometry-based proteomic analysis requires peptide fractionation to simplify complex biological samples and increase proteome coverage. OFFGEL fractionation technology became a common method to separate peptides or proteins using isoelectric focusing in an immobilized pH gradient. However, the OFFGEL focusing process may be further optimized and controlled in terms of separation time and pI resolution. Here, we evaluated OFFGEL technology to separate peptides from different samples in the presence of lowmolecular-weight (LMW) color pI markers to visualize the focusing process. LMW color pI markers covering a large pH range were added to the peptide mixture before OFFGEL fractionation using a 24-wells device encompassing the pH range 3-10. We also explored the impact of LMW color pI markers on peptide fractionation labeled previously for iTRAQ. Then, fractionated peptides were separated by RP_HPLC prior to MS analysis using MALDI-TOF/TOF mass spectrometry in MS and MS/MS modes. Here we report the performance of the peptide focusing process in the presence of LMW color pI markers as on-line trackers during the OFFGEL process and the possibility to use them as pI controls for peptide focusing. This method improves the workflow for peptide fractionation in a bottom-up proteomic approach with or without iTRAQ labeling.
PROTEOMICS, 2016
Besides their main contribution as substrates for protein synthesis, amino acids as signaling mol... more Besides their main contribution as substrates for protein synthesis, amino acids as signaling molecules could exert some regulatory functions on protein synthesis and/or proteolysis that have been emphasized in a number of recent studies. Several publications have highlighted supplemental roles of those amino acids in protein metabolism as well as in immunity, heat shock response, or apoptosis processes. In this way, via their regulatory properties, selected amino acids (such as leucine, glutamine, arginine, citrulline, or methionine) directly influence the proteome. In this review, we are proposing an overview of the regulation of the proteome by amino acids in mammals.