Structural and functional insights into Skl and Pal endolysins, two cysteine-amidases with anti-pneumococcal activity. Dithiothreitol (DTT) effect on lytic activity

20 pags, 9 figs, 2 tabs. -- The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmicb.2021.740914/full#supplementary-material

Bibliographic Details
Authors: Gallego-Páramo, Cristina, Hernández-Ortiz, Noelia, Buey, Rubén M., Rico-Lastres, Palma, García, Guadalupe, Díaz, José Fernando, García González, Pedro, Menéndez, Margarita
Format: article
Status:Published version
Publication Date:2021
Country:España
Institution:Consejo Superior de Investigaciones Científicas (CSIC)
Repository:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/258045
Online Access:http://hdl.handle.net/10261/258045
https://api.elsevier.com/content/abstract/scopus_id/85119078519
Access Level:Open access
Keyword:CHAP domain
DTT-mediated activation
Amidase_5 domain
Anti-pneumococcal activity
Choline-binding domain
Cysteine-peptidase
Endolysin
Reducing agents
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spelling Structural and functional insights into Skl and Pal endolysins, two cysteine-amidases with anti-pneumococcal activity. Dithiothreitol (DTT) effect on lytic activityGallego-Páramo, CristinaHernández-Ortiz, NoeliaBuey, Rubén M.Rico-Lastres, PalmaGarcía, GuadalupeDíaz, José FernandoGarcía González, PedroMenéndez, MargaritaCHAP domainDTT-mediated activationAmidase_5 domainAnti-pneumococcal activityCholine-binding domainCysteine-peptidaseEndolysinReducing agents20 pags, 9 figs, 2 tabs. -- The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmicb.2021.740914/full#supplementary-materialWe have structurally and functionally characterized Skl and Pal endolysins, the latter being the first endolysin shown to kill effectively Streptococcus pneumoniae, a leading cause of deathly diseases. We have proved that Skl and Pal are cysteine-amidases whose catalytic domains, from CHAP and Amidase_5 families, respectively, share an α3β6-fold with papain-like topology. Catalytic triads are identified (for the first time in Amidase_5 family), and residues relevant for substrate binding and catalysis inferred from in silico models, including a calcium-binding site accounting for Skl dependence on this cation for activity. Both endolysins contain a choline-binding domain (CBD) with a β-solenoid fold (homology modeled) and six conserved choline-binding loci whose saturation induced dimerization. Remarkably, Pal and Skl dimers display a common overall architecture, preserved in choline-bound dimers of pneumococcal lysins with other catalytic domains and bond specificities, as disclosed using small angle X-ray scattering (SAXS). Additionally, Skl is proved to be an efficient anti-pneumococcal agent that kills multi-resistant strains and clinical emergent-serotype isolates. Interestingly, Skl and Pal time-courses of pneumococcal lysis were sigmoidal, which might denote a limited access of both endolysins to target bonds at first stages of lysis. Furthermore, their DTT-mediated activation, of relevance for other cysteine-peptidases, cannot be solely ascribed to reversal of catalytic-cysteine oxidation.This work was supported by grants from the Ministry of Economy and Competitiveness (BFU2015-70072-R) and the Ministry of Science, Innovation and Universities (RTI2018-099985-B-I00/AEI/10.13039/501100011033) to MM, and by a grant from the Ministry of Economy and Competitiveness (MINECO-FEDER, SAF2017-88664-R) to PG. Additional funding was provided by the Centro de Investigacion Biomedica en Red de Enfermedades Respiratorias (CIBERES), an initiative of the Instituto de Salud Carlos III (ISCIII), to MM and PG.Peer reviewedFrontiers MediaMinisterio de Economía y Competitividad (España)Ministerio de Ciencia e Innovación (España)Agencia Estatal de Investigación (España)Centro de Investigación Biomédica en Red Enfermedades Respiratorias (España)Instituto de Salud Carlos IIIConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202220222021info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/258045https://api.elsevier.com/content/abstract/scopus_id/85119078519reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/MINECO//BFU2015-70072-Rinfo:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/SAF2017-88664-Rinfo:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/RTI2018-099985-B-I00Frontiers in microbiologyhttps://doi.org/10.3389/fmicb.2021.740914Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2580452026-05-22T06:33:51Z
dc.title.none.fl_str_mv Structural and functional insights into Skl and Pal endolysins, two cysteine-amidases with anti-pneumococcal activity. Dithiothreitol (DTT) effect on lytic activity
title Structural and functional insights into Skl and Pal endolysins, two cysteine-amidases with anti-pneumococcal activity. Dithiothreitol (DTT) effect on lytic activity
spellingShingle Structural and functional insights into Skl and Pal endolysins, two cysteine-amidases with anti-pneumococcal activity. Dithiothreitol (DTT) effect on lytic activity
Gallego-Páramo, Cristina
CHAP domain
DTT-mediated activation
Amidase_5 domain
Anti-pneumococcal activity
Choline-binding domain
Cysteine-peptidase
Endolysin
Reducing agents
title_short Structural and functional insights into Skl and Pal endolysins, two cysteine-amidases with anti-pneumococcal activity. Dithiothreitol (DTT) effect on lytic activity
title_full Structural and functional insights into Skl and Pal endolysins, two cysteine-amidases with anti-pneumococcal activity. Dithiothreitol (DTT) effect on lytic activity
title_fullStr Structural and functional insights into Skl and Pal endolysins, two cysteine-amidases with anti-pneumococcal activity. Dithiothreitol (DTT) effect on lytic activity
title_full_unstemmed Structural and functional insights into Skl and Pal endolysins, two cysteine-amidases with anti-pneumococcal activity. Dithiothreitol (DTT) effect on lytic activity
title_sort Structural and functional insights into Skl and Pal endolysins, two cysteine-amidases with anti-pneumococcal activity. Dithiothreitol (DTT) effect on lytic activity
dc.creator.none.fl_str_mv Gallego-Páramo, Cristina
Hernández-Ortiz, Noelia
Buey, Rubén M.
Rico-Lastres, Palma
García, Guadalupe
Díaz, José Fernando
García González, Pedro
Menéndez, Margarita
author Gallego-Páramo, Cristina
author_facet Gallego-Páramo, Cristina
Hernández-Ortiz, Noelia
Buey, Rubén M.
Rico-Lastres, Palma
García, Guadalupe
Díaz, José Fernando
García González, Pedro
Menéndez, Margarita
author_role author
author2 Hernández-Ortiz, Noelia
Buey, Rubén M.
Rico-Lastres, Palma
García, Guadalupe
Díaz, José Fernando
García González, Pedro
Menéndez, Margarita
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Economía y Competitividad (España)
Ministerio de Ciencia e Innovación (España)
Agencia Estatal de Investigación (España)
Centro de Investigación Biomédica en Red Enfermedades Respiratorias (España)
Instituto de Salud Carlos III
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv CHAP domain
DTT-mediated activation
Amidase_5 domain
Anti-pneumococcal activity
Choline-binding domain
Cysteine-peptidase
Endolysin
Reducing agents
topic CHAP domain
DTT-mediated activation
Amidase_5 domain
Anti-pneumococcal activity
Choline-binding domain
Cysteine-peptidase
Endolysin
Reducing agents
description 20 pags, 9 figs, 2 tabs. -- The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmicb.2021.740914/full#supplementary-material
publishDate 2021
dc.date.none.fl_str_mv 2021
2022
2022
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/258045
https://api.elsevier.com/content/abstract/scopus_id/85119078519
url http://hdl.handle.net/10261/258045
https://api.elsevier.com/content/abstract/scopus_id/85119078519
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/MINECO//BFU2015-70072-R
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/SAF2017-88664-R
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/RTI2018-099985-B-I00
Frontiers in microbiology
https://doi.org/10.3389/fmicb.2021.740914
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Frontiers Media
publisher.none.fl_str_mv Frontiers Media
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
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