المؤلفون: Merle Nicolas, S, Grunenwald, Anne, Figueres, Marie-Lucile, Chauvet, Sophie, Daugan, Marie, Knockaert, Samantha, Robe-Rybkine, Tania, Noe, Remi, May, Olivia, Frimat, Marie, Brinkman, Nathan, Gentinetta, Thomas, Miescher, Sylvia, Houillier, Pascal, Legros, Veronique, Gonnet, Florence, Blanc-Brude Olivier, P, Rabant, Marion, Daniel, Regis, Dimitrov Jordan, D, Roumenina Lubka, T

المساهمون: Inserm, Université de Lille, CHU Lille, Lille Inflammation Research International Center (LIRIC) - U995, Centre de Recherche des Cordeliers CRC (UMR_S_1138 / U1138), Lille Inflammation Research International Center - U 995 LIRIC, Université Paris-Saclay, Université Paris Descartes - Paris 5 UPD5

مصطلحات موضوعية: inflammation, endothelial activation, kidney injury, heme, hemolysis, experimental model of intravascular hemolysis, hemopexin, phenylhydrazine

الوصف: Intravascular erythrocyte destruction, accompanied by the release of pro-oxidative and pro-inflammatory components hemoglobin and heme, is a common event in the pathogenesis of numerous diseases with heterogeneous etiology and clinical features. A frequent adverse effect related to massive hemolysis is the renal injury and inflammation. Nevertheless, it is still unclear whether heme––a danger-associated molecular pattern––and ligand for TLR4 or upstream hemolysis-derived products are responsible for these effects. Well-characterized animal models of hemolysis with kidney impairment are needed to investigate how hemolysis drives kidney injury and to test novel therapeutic strategies. Here, we characterized the pathological processes leading to acute kidney injury and inflammation during massive intravascular hemolysis, using a mouse model of phenylhydrazine (PHZ)-triggered erythrocyte destruction. We observed profound changes in mRNA levels for markers of tubular damage (Kim-1, NGAL) and regeneration (indirect marker of tubular injury, Ki-67), and tissue and vascular inflammation (IL-6, E-selectin, P-selectin, ICAM-1) in kidneys of PHZ-treated mice, associated with ultrastructural signs of tubular injury. Moreover, mass spectrometry revealed presence of markers of tubular damage in urine, including meprin-α, cytoskeletal keratins, α-1-antitrypsin, and α-1-microglobulin. Signs of renal injury and inflammation rapidly resolved and the renal function was preserved, despite major changes in metabolic parameters of PHZ-injected animals. Mechanistically, renal alterations were largely heme-independent, since injection of free heme could not reproduce them, and scavenging heme with hemopexin in PHZ-administered mice could not prevent them. Reduced overall health status of the mice suggested multiorgan involvement. We detected amylasemia and amylasuria, two markers of acute pancreatitis. We also provide detailed characterization of renal manifestations associated with acute intravascular hemolysis, which may be mediated ...

وصف الملف: application/octet-stream

العلاقة: Frontiers in Immunology; Front. Immunol.; http://hdl.handle.net/20.500.12210/5080Test

الإتاحة: https://doi.org/20.500.12210/5080Test
https://hdl.handle.net/20.500.12210/5080Test

المؤلفون: Seffouh, Ilham, Bilong, Mélanie, Przybylski, Cédric, El Omrani, Nesrine, Poyer, Salomé, Lamour, Guillaume, Clément, Marie-Jeanne, Boustany, Rebecca-Joe, Gout, Evelyne, Gonnet, Florence, Vivès, Romain, R, Daniel, Régis

المساهمون: Université Paris-Saclay, Institut de biologie structurale (IBS - UMR 5075), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA), Centre National de la Recherche Scientifique (CNRS), Laboratoire Analyse, Modélisation et Matériaux pour la Biologie et l'Environnement (LAMBE - UMR 8587), Université d'Évry-Val-d'Essonne (UEVE)-Institut de Chimie - CNRS Chimie (INC-CNRS)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-CY Cergy Paris Université (CY)

المصدر: ISSN: 2045-2322.

مصطلحات موضوعية: [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [CHIM.ANAL]Chemical Sciences/Analytical chemistry, [SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM]

الوصف: International audience ; AbstractThe human sulfatase HSulf-2 is one of only two known endosulfatases that play a decisive role in modulating the binding properties of heparan sulfate proteoglycans on the cell surface and in the extracellular matrix. Recently, HSulf-2 was shown to exhibit an unusual post-translational modification consisting of a sulfated glycosaminoglycan chain. This study describes the structural characterization of this glycosaminoglycan (GAG) and provides new data on its impact on the catalytic properties of HSulf-2. The unrevealed nature of this GAG chain is identified as a chondroitin/dermatan sulfate (CS/DS) mixed chain, as shown by mass spectrometry combined with NMR analysis. It consists primarily of 6-O and 4-O monosulfated disaccharide units, with a slight predominance of the 4-O-sulfation. Using atomic force microscopy, we show that this unique post-translational modification dramatically impacts the enzyme hydrodynamic volume. We identified human hyaluronidase-4 as a secreted hydrolase that can digest HSulf-2 GAG chain. We also showed that HSulf-2 is able to efficiently 6-O-desulfate antithrombin III binding pentasaccharide motif, and that this activity was enhanced upon removal of the GAG chain. Finally, we identified five N-glycosylation sites on the protein and showed that, although required, reduced N-glycosylation profiles were sufficient to sustain HSulf-2 integrity.

العلاقة: hal-04348199; https://hal.science/hal-04348199Test; https://hal.science/hal-04348199/documentTest; https://hal.science/hal-04348199/file/Seffouh_et_al-2023-Scientific_Reports.pdfTest

الإتاحة: https://doi.org/10.1038/s41598-023-49147-5Test
https://hal.science/hal-04348199Test
https://hal.science/hal-04348199/documentTest
https://hal.science/hal-04348199/file/Seffouh_et_al-2023-Scientific_Reports.pdfTest

المؤلفون: El Omrani, Nesrine, Poyer, Salomé, Takeda-Uchimura, Yoshiko, Uchimura, Kenji, Vivès, Romain R, Gonnet, Florence, Daniel, Régis

المساهمون: Laboratoire Analyse, Modélisation et Matériaux pour la Biologie et l'Environnement (LAMBE - UMR 8587), Université d'Évry-Val-d'Essonne (UEVE)-Institut de Chimie - CNRS Chimie (INC-CNRS)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-CY Cergy Paris Université (CY), Institut de Chimie des Substances Naturelles (ICSN), Institut de Chimie - CNRS Chimie (INC-CNRS)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institut de biologie structurale (IBS - UMR 5075 ), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes 2016-2019 (UGA 2016-2019 )-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Centre National de la Recherche Scientifique (CNRS), Université Paris-Saclay

المصدر: IMMS 2023 ; https://hal.science/hal-04428436Test ; IMMS 2023, May 2023, Paris, France

مصطلحات موضوعية: [CHIM]Chemical Sciences

جغرافية الموضوع: Paris, France

الوصف: International audience ; Glycosaminoglycans (GAGs) are anionic polysaccharides of remarkable molecular complexity involved invarious biological and physio-pathological processes. The determination of structure-functionrelationships among these molecules is of great interest; however, the complex structure of GAGs, ofwhich heparan sulfate (HS) is the most challenging representative, and the lack of tools for decipheringcomplex GAG sequences has restricted advances in the GAGs field. In fact, at the molecular level, HSconstitutive disaccharide units can be modified by acetylation, epimerization, and sulfation at multiplepositions by highly regulated biosynthetic machinery. These modifications are completed by a post-synthetic editing process involving endosulfatases that finely tune the sulfate code along the HS chain.In humans, HSulf-1 and HSulf-2 are extracellular sulfatases that regioselectively remove the 6-O-sulfategroups from HS. HSulfs action alters HS ligand binding properties and modulates multiple signalingpathways. To gain new insights into the functional properties of HSulf enzymes, we set up a robust andresolving analytical method based on hydrophilic interaction liquid chromatography (HILIC) coupled withmass spectrometry (MS). This method allowed the structural determination of the enzyme productsfrom various sulfated oligosaccharide substrates and the monitoring of the 6-O-sulfate hydrolysis ofnatural sulfated substrates by HSulf enzymes. HILIC-MS methods are developed in our laboratory toallow the separation of GAG-sulfated oligosaccharides by size and sulfate patern. A specificmethodology was developed to monitor the progress of the enzyme reaction catalyzed by theendosulfatase HSulfs on various heparin (Hp)-derived oligosaccharides and characterize both thestructure and the kinetics of the formation of the enzyme products. We followed the desulfation reactionon various heparin-oligosaccharide substrates over time. The reaction conditions of the heparinoligosaccharide substrates with the enzymes ...

العلاقة: hal-04428436; https://hal.science/hal-04428436Test

الإتاحة: https://hal.science/hal-04428436Test

المؤلفون: El Omrani, Nesrine, Poyer, Salomé, Przybylski, Cédric, Takeda-Uchimura, Yoshiko, Uchimura, Kenji, Vivès, Romain R, Gonnet, Florence, Daniel, Régis

المساهمون: Laboratoire Analyse, Modélisation et Matériaux pour la Biologie et l'Environnement (LAMBE - UMR 8587), Université d'Évry-Val-d'Essonne (UEVE)-Institut de Chimie - CNRS Chimie (INC-CNRS)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-CY Cergy Paris Université (CY), Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576 (UGSF), Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institut de biologie structurale (IBS - UMR 5075 ), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes 2016-2019 (UGA 2016-2019 )-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Centre National de la Recherche Scientifique (CNRS), Université Paris-Saclay

المصدر: IMMS 2023 ; https://hal.science/hal-04429176Test ; IMMS 2023, Jul 2023, Paris, France

مصطلحات موضوعية: [CHIM.ANAL]Chemical Sciences/Analytical chemistry

جغرافية الموضوع: Paris, France

الوصف: International audience ; Glycosaminoglycans (GAGs) are anionic polysaccharides of remarkable molecular complexity involved in various biological and physio-pathological processes. The determination of structure-function relationships among these molecules is of great interest; however, the complex structure of GAGs, of which heparan sulfate (HS) is the most challenging representative, and the lack of tools for deciphering complex GAG sequences has restricted advances in the GAGs field. In fact, at the molecular level, HS constitutive disaccharide units can be modified by acetylation, epimerization, and sulfation at multiple positions by highly regulated biosynthetic machinery. These modifications are completed by a post-synthetic editing process involving endosulfatases that finely tune the sulfate code along the HS chain.In humans, HSulf-1 and HSulf-2 are extracellular sulfatases that regioselectively remove the 6-O-sulfate groups from HS. HSulfs action alters HS ligand binding properties and modulates multiple signaling pathways. To gain new insights into the functional properties of HSulf enzymes, we set up a robust and resolving analytical method based on hydrophilic interaction liquid chromatography (HILIC) coupled with mass spectrometry (MS). This method allowed the structural determination of the enzyme products from various sulfated oligosaccharide substrates and the monitoring of the 6-O-sulfate hydrolysis of natural sulfated substrates by HSulf enzymes. HILIC-MS methods are developed in our laboratory to allow the separation of GAG-sulfated oligosaccharides by size and sulfate patern. A specific methodology was developed to monitor the progress of the enzyme reaction catalyzed by the endosulfatase HSulfs on various heparin (Hp)-derived oligosaccharides and characterize both the structure and the kinetics of the formation of the enzyme products. We followed the desulfation reaction on various heparin-oligosaccharide substrates over time. The reaction conditions of the heparin oligosaccharide substrates ...

العلاقة: hal-04429176; https://hal.science/hal-04429176Test

الإتاحة: https://hal.science/hal-04429176Test

المؤلفون: Przybylski, Cédric, Gonnet, Florence, Saesen, Els, Lortat-Jacob, Hugues, Daniel, Régis

المساهمون: Laboratoire Analyse, Modélisation et Matériaux pour la Biologie et l'Environnement (LAMBE - UMR 8587), Université d'Évry-Val-d'Essonne (UEVE)-Institut de Chimie - CNRS Chimie (INC-CNRS)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS)-CY Cergy Paris Université (CY), Institut Parisien de Chimie Moléculaire (IPCM), Chimie Moléculaire de Paris Centre (FR 2769), École normale supérieure - Paris (ENS-PSL), Université Paris Sciences et Lettres (PSL)-Université Paris Sciences et Lettres (PSL)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris Sciences et Lettres (PSL)-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris Sciences et Lettres (PSL)-Institut de Chimie - CNRS Chimie (INC-CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris Sciences et Lettres (PSL)-Institut de Chimie - CNRS Chimie (INC-CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut de biologie structurale (IBS - UMR 5075), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)

المصدر: ISSN: 1618-2642.

مصطلحات موضوعية: [CHIM.ANAL]Chemical Sciences/Analytical chemistry

الوصف: International audience

العلاقة: hal-02401458; https://univ-evry.hal.science/hal-02401458Test; https://univ-evry.hal.science/hal-02401458/documentTest; https://univ-evry.hal.science/hal-02401458/file/Przybylski%20et%20al.%20-%202019%20-%20Surface%20plasmon%20resonance%20imaging%20coupled%20to%20on-ch.pdfTest

الإتاحة: https://doi.org/10.1007/s00216-019-02267-2Test
https://univ-evry.hal.science/hal-02401458Test
https://univ-evry.hal.science/hal-02401458/documentTest
https://univ-evry.hal.science/hal-02401458/file/Przybylski%20et%20al.%20-%202019%20-%20Surface%20plasmon%20resonance%20imaging%20coupled%20to%20on-ch.pdfTest

المؤلفون: Seffouh, Ilham, Przybylski, Cédric, Seffouh, Amal, El Masri, Rana, Vivès, Romain R, Gonnet, Florence, Daniel, Régis

المساهمون: Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement (LAMBE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine-Université d'Évry-Val-d'Essonne (UEVE)-Institut de Chimie - CNRS Chimie (INC-CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Saclay (COmUE), Institut de biologie structurale (IBS - UMR 5075 ), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes 2016-2019 (UGA 2016-2019 )-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), ANR-17-CE11-0040,SULFatAS,Structure et activités des sulfatases extracellulaires SULF(2017)

المصدر: ISSN: 2405-5808 ; Biochemistry and Biophysics Reports ; https://hal.science/hal-02043614Test ; Biochemistry and Biophysics Reports, 2019, 18, pp.100617. ⟨10.1016/j.bbrep.2019.01.010⟩.

مصطلحات موضوعية: HSulf-2, Heparan sulfate, Mass spectrometry, Sulfatase, 6-O-Endosulfatase, Formylglycine, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM]

الوصف: International audience ; The human 6-O-endosulfatases HSulf-1 and -2 catalyze the region-selective hydrolysis of the 6-O-sulfate group of the glucosamine residues within sulfated domains of heparan sulfate, thereby ensuring a unique and original post-biosynthetic modification of the cell surface proteoglycans. While numerous studies point out the role of HSulf-2 in crucial physiological processes as well as in pathological conditions particularly in cancer, its structural organization in two chains and its functional properties remain poorly understood. In this study, we report the first characterization by mass spectrometry (MS) of HSulf-2. An average molecular weight of 133,115 Da was determined for the whole enzyme by MALDI-TOF MS, i.e. higher than the naked amino acid backbone (98,170 Da), highlighting a significant contribution of post-translational modifications. The HSulf-2 protein sequence was determined by Nano-LC-MS/MS, leading to 63% coverage and indicating at least four N-glycosylation sites at Asn 108, 147, 174 and 217. These results provide a platform for further structural investigations of the HSulf enzymes, aiming at deciphering the role of each chain in the substrate binding and specificities and in the catalytic activities.

العلاقة: info:eu-repo/semantics/altIdentifier/pmid/30788440; hal-02043614; https://hal.science/hal-02043614Test; https://hal.science/hal-02043614/documentTest; https://hal.science/hal-02043614/file/main.pdfTest; PUBMED: 30788440; PUBMEDCENTRAL: PMC6369371

الإتاحة: https://doi.org/10.1016/j.bbrep.2019.01.010Test
https://hal.science/hal-02043614Test
https://hal.science/hal-02043614/documentTest
https://hal.science/hal-02043614/file/main.pdfTest

المؤلفون: Merle, Nicolas, Grunenwald, Anne, Figueres, Marie-Lucile, Chauvet, Sophie, Daugan, Marie, Knockaert, Samantha, Robe-Rybkine, Tania, Noe, Remi, May, Olivia, Frimat, Marie, Brinkman, Nathan, Gentinetta, Thomas, Miescher, Sylvia, Houillier, Pascal, Legros, Véronique, Gonnet, Florence, Blanc-Brude, Olivier, Rabant, Marion, Daniel, Régis, Dimitrov, Jordan, Roumenina, Lubka

المساهمون: Centre de Recherche des Cordeliers (CRC (UMR_S_1138 / U1138)), École Pratique des Hautes Études (EPHE), Université Paris Sciences et Lettres (PSL)-Université Paris Sciences et Lettres (PSL)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Hôpital Européen Georges Pompidou APHP (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO), Lille Inflammation Research International Center - U 995 (LIRIC), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire CHU Lille (CHRU Lille), CSL Behring, Service de Physiologie Georges-Pompidou, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Européen Georges Pompidou APHP (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Université Paris Descartes - Paris 5 (UPD5), Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement (LAMBE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine-Université d'Évry-Val-d'Essonne (UEVE)-Institut de Chimie - CNRS Chimie (INC-CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Saclay (COmUE), Paris-Centre de Recherche Cardiovasculaire (PARCC - UMR-S U970), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Hôpital Necker - Enfants Malades AP-HP, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), ANR-15-CE15-0001,INFLACOMP,Inflammation et lésions endothéliales induites par le Complément et l'hémolyse(2015), ANR-12-JSV1-0006,PIKinART,Role de la phosphoinositide 3-kinase dans les pathologies artérielles: une cible pharmacologique prometteuse(2012), ANR-11-IDEX-0005,USPC,Université Sorbonne Paris Cité(2011)

المصدر: ISSN: 1664-3224.

مصطلحات موضوعية: hemolysis, heme, kidney injury, endothelial activation, inflammation, phenylhydrazine, experimental model of intravascular hemolysis, hemopexin, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, [SDV.MHEP.UN]Life Sciences [q-bio]/Human health and pathology/Urology and Nephrology

الوصف: International audience ; Intravascular erythrocyte destruction, accompanied by the release of pro-oxidative and pro-inflammatory components hemoglobin and heme, is a common event in the pathogenesis of numerous diseases with heterogeneous etiology and clinical features. A frequent adverse effect related to massive hemolysis is the renal injury and inflammation. Nevertheless, it is still unclear whether heme––a danger-associated molecular pattern––and ligand for TLR4 or upstream hemolysis-derived products are responsible for these effects. Well-characterized animal models of hemolysis with kidney impairment are needed to investigate how hemolysis drives kidney injury and to test novel therapeutic strategies. Here, we characterized the pathological processes leading to acute kidney injury and inflammation during massive intravascular hemolysis, using a mouse model of phenylhydrazine (PHZ)-triggered erythrocyte destruction. We observed profound changes in mRNA levels for markers of tubular damage (Kim-1, NGAL) and regeneration (indirect marker of tubular injury, Ki-67), and tissue and vascular inflammation (IL-6, E-selectin, P-selectin, ICAM-1) in kidneys of PHZ-treated mice, associated with ultrastructural signs of tubular injury. Moreover, mass spectrometry revealed presence of markers of tubular damage in urine, including meprin-α, cytoskeletal keratins, α-1-antitrypsin, and α-1-microglobulin. Signs of renal injury and inflammation rapidly resolved and the renal function was preserved, despite major changes in metabolic parameters of PHZ-injected animals. Mechanistically, renal alterations were largely heme-independent, since injection of free heme could not reproduce them, and scavenging heme with hemopexin in PHZ-administered mice could not prevent them. Reduced overall health status of the mice suggested multiorgan involvement. We detected amylasemia and amylasuria, two markers of acute pancreatitis. We also provide detailed characterization of renal manifestations associated with acute intravascular ...

العلاقة: hal-01966724; https://hal.science/hal-01966724Test; https://hal.science/hal-01966724/documentTest; https://hal.science/hal-01966724/file/fimmu-09-00179.pdfTest

الإتاحة: https://doi.org/10.3389/fimmu.2018.00179Test
https://hal.science/hal-01966724Test
https://hal.science/hal-01966724/documentTest
https://hal.science/hal-01966724/file/fimmu-09-00179.pdfTest

المؤلفون: Baud, Anna, Aymé, Laure, Gonnet, Florence, Salard, Isabelle, Gohon, Yann, Jolivet, Pascale, Brodolin, Konstantin, da Silva, P., Giuliani, Alexandre, Sclavi, B., Chardot, Thierry, Mercere, P., Roblin, Pierre, Daniel, Régis

المساهمون: Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement (LAMBE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine-Université d'Évry-Val-d'Essonne (UEVE)-Institut de Chimie - CNRS Chimie (INC-CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Saclay (COmUE), Institut Jean-Pierre Bourgin (IJPB), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Centre d’études d’Agents Pathogènes et Biotechologies pour la Santé (CPBS), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Synchrotron SOLEIL (SSOLEIL), Centre National de la Recherche Scientifique (CNRS), Département Caractérisation et Elaboration des Produits Issus de l'Agriculture (CEPIA), Institut National de la Recherche Agronomique (INRA), Laboratoire de Biologie et de Pharmacologie Appliquée (LBPA), École normale supérieure - Cachan (ENS Cachan)-Centre National de la Recherche Scientifique (CNRS), SOLEIL 20120893, 20130817, 20140760, INRA CEPIA department: OlSoleilMiox project

المصدر: ISSN: 0909-0495.

مصطلحات موضوعية: factor H, C3b, mass spectrometry, X-ray footprinting, structural proteomics, S3 oleosin, [SDV]Life Sciences [q-bio], [CHIM.ANAL]Chemical Sciences/Analytical chemistry, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM]

الوصف: Synchrotron X-ray footprinting complements the techniques commonly used to define the structure of molecules such as crystallography, small-angle X-ray scattering and nuclear magnetic resonance. It is remarkably useful in probing the structure and interactions of proteins with lipids, nucleic acids or with other proteins in solution, often better reflecting the in vivo state dynamics. To date, most X-ray footprinting studies have been carried out at the National Synchrotron Light Source, USA, and at the European Synchrotron Radiation Facility in Grenoble, France. This work presents X-ray footprinting of biomolecules performed for the first time at the X-ray Metrology beamline at the SOLEIL synchrotron radiation source. The installation at this beamline of a stopped-flow apparatus for sample delivery, an irradiation capillary and an automatic sample collector enabled the X-ray footprinting study of the structure of the soluble protein factor H (FH) from the human complement system as well as of the lipid-associated hydrophobic protein S3 oleosin from plant seed. Mass spectrometry analysis showed that the structural integrity of both proteins was not affected by the short exposition to the oxygen radicals produced during the irradiation. Irradiated molecules were subsequently analysed using high-resolution mass spectrometry to identify and locate oxidized amino acids. Moreover, the analyses of FH in its free state and in complex with complement C3b protein have allowed us to create a map of reactive solvent-exposed residues on the surface of FH and to observe the changes in oxidation of FH residues upon C3b binding. Studies of the solvent accessibility of the S3 oleosin show that X-ray footprinting offers also a unique approach to studying the structure of proteins embedded within membranes or lipid bodies. All the biomolecular applications reported herein demonstrate that the Metrology beamline at SOLEIL can be successfully used for synchrotron X-ray footprinting of biomolecules.

العلاقة: info:eu-repo/semantics/altIdentifier/pmid/28452748; hal-01608337; https://hal.science/hal-01608337Test; https://hal.science/hal-01608337/documentTest; https://hal.science/hal-01608337/file/2017_Baud_J.%20Synchrotron%20Rad_1.pdfTest; PRODINRA: 398098; PUBMED: 28452748; WOS: 000400385400004

الإتاحة: https://doi.org/10.1107/S1600577517002478Test
https://hal.science/hal-01608337Test
https://hal.science/hal-01608337/documentTest
https://hal.science/hal-01608337/file/2017_Baud_J.%20Synchrotron%20Rad_1.pdfTest

المؤلفون: Bally, Isabelle, Ancelet, Sarah, Moriscot, Christine, Gonnet, Florence, Mantovani, Alberto, Daniel, Régis, Schoehn, Guy, Arlaud, Gérard J., Thielens, Nicole M.

المصدر: Proceedings of the National Academy of Sciences of the United States of America, 2013 May . 110(21), 8650-8655.

الوصول الحر: https://www.jstor.org/stable/42656777Test

المؤلفون: Baud, Anna, Gonnet, Florence, Salard, Isabelle, Le Mignon, Maxime, Giuliani, Alexandre, Mercère, Pascal, Sclavi, Bianca, Daniel, Régis

المساهمون: Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement (LAMBE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine-Université d'Évry-Val-d'Essonne (UEVE)-Institut de Chimie - CNRS Chimie (INC-CNRS)-Centre National de la Recherche Scientifique (CNRS), Synchrotron SOLEIL (SSOLEIL), Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biologie et de Pharmacologie Appliquée (LBPA), École normale supérieure - Cachan (ENS Cachan)-Centre National de la Recherche Scientifique (CNRS), the CNRS, the Evry-Val-d'Essonne University, the Genopole

المصدر: ISSN: 0264-6021.

مصطلحات موضوعية: complement system, Factor H, mass spectrometry, hydroxyl radical protein footprinting, C3b protein, cross-linking, [SDV.IMM.II]Life Sciences [q-bio]/Immunology/Innate immunity, [CHIM.ANAL]Chemical Sciences/Analytical chemistry

الوصف: International audience ; The control protein Factor H (FH) is a crucial regulator of the innate immune complement system, where it is active on host cell membranes and in the fluid phase. Mutations impairing the binding capacity of FH lead to severe autoimmune diseases. Here, we studied the solution structure of full-length FH, in its free state and bound to the C3b complement protein. To do so, we used two powerful techniques, hydroxyl radical protein footprinting (HRPF) and chemical cross-linking coupled with mass spectrometry (MS), to probe the structural rearrangements and to identify protein interfaces. The footprint of C3b on the FH surface matches existing crystal structures of C3b complexed with the N- and C-terminal fragments of FH. In addition, we revealed the position of the central portion of FH in the protein complex. Moreover, cross-linking studies confirmed the involvement of the C-terminus in the dimerization of FH.

العلاقة: info:eu-repo/semantics/altIdentifier/pmid/27099340; hal-01966773; https://hal.science/hal-01966773Test; https://hal.science/hal-01966773/documentTest; https://hal.science/hal-01966773/file/Baud%20et%20al%202016%20Probing%20the%20solution%20structure%20of%20Factor%20H%20using%20hydroxyl%20radical%20protein%20footprinting%20and%20cross-linking.pdfTest; PUBMED: 27099340

الإتاحة: https://doi.org/10.1042/BCJ20160225Test
https://hal.science/hal-01966773Test
https://hal.science/hal-01966773/documentTest
https://hal.science/hal-01966773/file/Baud%20et%20al%202016%20Probing%20the%20solution%20structure%20of%20Factor%20H%20using%20hydroxyl%20radical%20protein%20footprinting%20and%20cross-linking.pdfTest