High-Resolution Mass Spectrometry-Based Approaches for the Detection and Quantification of Peptidase Activity in Plasma
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Le résumé fourni par la source
Proteomic technologies have identified 234 peptidases in plasma but little quantitative information about the proteolytic activity has been uncovered. In this study, the substrate profile of plasma proteases was evaluated using two nano-LC-ESI-MS/MS methods. Multiplex substrate profiling by mass spectrometry (MSP-MS) quantifies plasma protease activity in vitro using a global and unbiased library of synthetic peptide reporter substrates, and shotgun peptidomics quantifies protein degradation products that have been generated in vivo by proteases. The two approaches gave complementary results since they both highlight key peptidase activities in plasma including amino- and carboxypeptidases with different substrate specificity profiles. These assays provide a significant advantage over traditional approaches, such as fluorogenic peptide reporter substrates, because they can detect active plasma proteases in a global and unbiased manner, in comparison to detecting select proteases using specific reporter substrates. We discovered that plasma proteins are cleaved by endoproteases and these peptide products are subsequently degraded by amino- and carboxypeptidases. The exopeptidases are more active and stable in plasma and therefore were found to be the most active proteases in the in vitro assay. The protocols presented here set the groundwork for studies to evaluate changes in plasma proteolytic activity in shock.
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Le contrôle bibliographique ouvert
DOI retrouvé dans Crossref DOI retrouvé ; titre concordant.
- Titre Crossref
- High-Resolution Mass Spectrometry-Based Approaches for the Detection and Quantification of Peptidase Activity in Plasma
- Date Crossref
- 06/09/2020
- Éditeur
- MDPI AG
- Type
- journal-article
Ce recoupement confirme des métadonnées liées au DOI. Il ne confirme ni la méthode ni les conclusions de l’étude, et il ne compte pas comme une seconde source scientifique indépendante.
Où se fait cette recherche
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University of Milan Centre for Nanostructured Materials and Interfaces (CIMAINA) pays non établi dans la noticeUniversité ou école supérieure
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University of California San Diego Department of Anesthesiology & pays non établi dans la noticeUniversité ou école supérieure
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Politecnico di Milano pays non établi dans la noticeUniversité ou école supérieure
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VA San Diego Healthcare System pays non établi dans la noticeÉtablissement de santé
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Skaggs School of Pharmacy and Pharmaceutical Sciences pays non établi dans la noticeUniversité ou école supérieure
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School of Medicine Department of Neurosciences pays non établi dans la noticeUniversité ou école supérieure
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Dipartimento di Elettronica pays non établi dans la noticeInstitution
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Department of Anesthesiology & pays non établi dans la noticeInstitution
Centre for Nanostructured Materials and Interfaces (CIMAINA) — University of Milan, Department of Anesthesiology & — University of California San Diego et Politecnico di Milano, avec 5 autres affiliations.
Une affiliation ne permet pas de déduire la nationalité d’un auteur.