Mostrar el registro sencillo del ítem
dc.contributor.author
Covaleda, Giovanni
dc.contributor.author
Trejo, Sebastian Alejandro
dc.contributor.author
Salas Sarduy, Emir
dc.contributor.author
del Rivero, Maday Alonso
dc.contributor.author
Chavez, Maria Angeles
dc.contributor.author
Aviles, Francesc X.
dc.date.available
2018-12-04T17:15:30Z
dc.date.issued
2017-08
dc.identifier.citation
Covaleda, Giovanni; Trejo, Sebastian Alejandro; Salas Sarduy, Emir; del Rivero, Maday Alonso; Chavez, Maria Angeles; et al.; Intensity fading MALDI-TOF mass spectrometry and functional proteomics assignments to identify protease inhibitors in marine invertebrates; Elsevier Science; Journal Of Proteomics; 165; 8-2017; 75-92
dc.identifier.issn
1874-3919
dc.identifier.uri
http://hdl.handle.net/11336/65724
dc.description.abstract
Proteases and their inhibitors have become molecules of increasing fundamental and applicative value. Here we report an integrated strategy to identify and analyze such inhibitors from Caribbean marine invertebrates extracts by a fast and sensitive functional proteomics-like approach. The strategy works in three steps: i) multiplexed enzymatic inhibition kinetic assays, ii) Intensity Fading MALDI-TOF MS to establish a link between inhibitory molecules and the related MALDI signal(s) detected in the extract(s), and iii) ISD-CID-T3 MS fragmentation on the parent MALDI signals selected in the previous step, enabling the partial or total top-down sequencing of the molecules. The present study has allowed validation of the whole approach, identification of a substantial number of novel protein protease inhibitors, as well as full or partial sequencing of reference molecular species and of many unknown ones, respectively. Such inhibitors correspond to six protease subfamilies (metallocarboxypeptidases-A and -B, pepsin, papain, trypsin and subtilisin), are small (1–10 KDa) disulfide-rich proteins, and have been found at diverse frequencies among the invertebrates (13 to 41%). The overall procedure could be tailored to other enzyme-inhibitor and protein interacting systems, analyzing samples at medium-throughput level and leading to the functional and structural characterization of proteinaceous ligands from complex biological extracts. Significance Invertebrate animals, and marine ones among, display a remarkable diversity of species and contained biomolecules. Many of their proteins-peptides have high biological, biotechnological and biomedical potential interest but, because of the lack of sequenced genomes behind, their structural and functional characterization constitutes a great challenge. Here, looking at the small, disulfide-rich, proteinaceous inhibitors of proteases found in them, it is shown that such problem can be significatively facilitated by integrative multiplexed enzymatic assays, affinity-based Intensity-Fading (IF-) MALDI-TOF mass spectrometry (MS), and on-line MS fragmentation, in a fast and easy approach.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Elsevier Science
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Complex Biological Samples
dc.subject
Invertebrate
dc.subject
Mass Spectrometry
dc.subject
Medium-Throughput Screening
dc.subject
Protease
dc.subject
Protease Inhibitor
dc.subject
Proteomics
dc.subject.classification
Otras Ciencias Biológicas
dc.subject.classification
Ciencias Biológicas
dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
dc.title
Intensity fading MALDI-TOF mass spectrometry and functional proteomics assignments to identify protease inhibitors in marine invertebrates
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2018-10-22T21:50:32Z
dc.journal.volume
165
dc.journal.pagination
75-92
dc.journal.pais
Países Bajos
dc.journal.ciudad
Amsterdam
dc.description.fil
Fil: Covaleda, Giovanni. Universitat Autònoma de Barcelona; España
dc.description.fil
Fil: Trejo, Sebastian Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universitat Autònoma de Barcelona; España
dc.description.fil
Fil: Salas Sarduy, Emir. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de la Habana; Cuba
dc.description.fil
Fil: del Rivero, Maday Alonso. Universidad de la Habana; Cuba
dc.description.fil
Fil: Chavez, Maria Angeles. Universidad de la Habana; Cuba
dc.description.fil
Fil: Aviles, Francesc X.. Universitat Autònoma de Barcelona; España
dc.journal.title
Journal Of Proteomics
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://linkinghub.elsevier.com/retrieve/pii/S1874391917301963
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/https://dx.doi.org/10.1016/j.jprot.2017.05.027
Archivos asociados