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Pre-miRNA-149 G-quadruplex as a molecular agent to capture nucleolin

2022-02-01Artigo científico Acesso aberto
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dc.contributor.authorSantos, Tiago
dc.contributor.authorMiranda, André
dc.contributor.authorImbert, Lionel
dc.contributor.authorJardim, Andreia
dc.contributor.authorCaneira, Catarina R. F.
dc.contributor.authorChu, Virgínia
dc.contributor.authorConde, João Pedro
dc.contributor.authorCampello, Maria Paula Cabral
dc.contributor.authorPaulo, António
dc.contributor.authorSalgado, Gilmar
dc.contributor.authorCabrita, Eurico J.
dc.contributor.authorCruz, Carla
dc.contributor.institutionUCIBIO - Applied Molecular Biosciences Unit
dc.contributor.institutionDQ - Departamento de Química
dc.contributor.pblElsevier BV
dc.date.accessioned2022-03-11T23:22:16Z
dc.date.available2022-03-11T23:22:16Z
dc.date.issued2022-02-01
dc.descriptionPD/BD/142851/2018 PD/00065/2013 MIT-EXPL/BIO/0008/2017 IF/00959/2015
dc.description.abstractOne of the most significant challenges in capturing and detecting biomarkers is the choice of an appropriate biomolecular receptor. Recently, RNA G-quadruplexes emerged as plausible receptors due to their ability to recognize with high-affinity proteins. Herein, we have unveiled and characterized the capability of the precursor microRNA 149 to form a G-quadruplex structure and determined the role that some ligands may have in its folding and binding capacity to nucleolin. The G-quadruplex formation was induced by K+ ions and stabilized by ligands, as demonstrated by nuclear magnetic resonance and circular dichroism experiments. Surface plasmon resonance measurements showed a binding affinity of precursor microRNA 149 towards ligands in the micromolar range (10−5–10−6 M) and a strong binding affinity to nucleolin RNA-binding domains 1 and 2 (8.38 × 10−10 M). Even in the presence of the ligand PhenDC3, the binding remains almost identical and in the same order of magnitude (4.46 × 10−10 M). The molecular interactions of the RNA G-quadruplex motif found in precursor miRNA 149 (5′-GGGAGGGAGGGACGGG- 3′) and nucleolin RNA-binding domains 1 and 2 were explored by means of molecular docking and molecular dynamics studies. The results showed that RNA G-quadruplex binds to a cavity between domains 1 and 2 of the protein. Then, complex formation was also evaluated through polyacrylamide gel electrophoresis. The results suggest that precursor microRNA 149/ligands and precursor microRNA 149/nucleolin RNA-binding domains 1 and 2 form stable molecular complexes. The in vitro co-localization of precursor microRNA 149 and nucleolin in PC3 cells was demonstrated using confocal microscopy. Finally, a rapid and straightforward microfluidic strategy was employed to check the ability of precursor microRNA 149 to capture nucleolin RNA-binding domains 1 and 2. The results revealed that precursor microRNA 149 can capture nucleolin RNA-binding domains 1 and 2 labeled with Fluorescein 5-isothiocyanate in a concentration-dependent manner, but PhenDC3 complexation seems to decrease the ability of precursor microRNA 149 to capture the protein. Overall, our results proved the formation of the G-quadruplex structure in the precursor microRNA 149 and the ability to recognize and detect nucleolin. This proof-of-concept study could open up a new framework for developing new strategies to design improved molecular receptors for capture and detection of nucleolin in complex biological samples.en
dc.description.versionpublishersversion
dc.description.versionpublished
dc.format.extent7075072
dc.identifier.doi10.1016/j.ejps.2021.106093
dc.identifier.issn0928-0987
dc.identifier.otherPURE: 36467709
dc.identifier.otherPURE UUID: ad19b4dc-2f05-4995-a7ec-cdc3fc9664e9
dc.identifier.otherScopus: 85121779205
dc.identifier.otherPubMed: 34922315
dc.identifier.otherWOS: 000804547300006
dc.identifier.urihttp://hdl.handle.net/10362/134341
dc.identifier.urlhttps://www.scopus.com/pages/publications/85121779205
dc.language.isoeng
dc.peerreviewedyes
dc.relationinfo:eu-repo/grantAgreement/EC/FP7/205079/EU
dc.relationUnlocking Animal Food Quality Research Potential in Baltic Region by Developing Scientific and Technical Capacities of the Research Institute “Sigra”
dc.subjectG-quadruplex ligands
dc.subjectMolecular receptors
dc.subjectNucleolin
dc.subjectPre-miRNA-149
dc.subjectRNA G-quadruplex
dc.subjectPharmaceutical Science
dc.titlePre-miRNA-149 G-quadruplex as a molecular agent to capture nucleolinen
dc.typejournal article
degois.publication.titleEuropean Journal Of Pharmaceutical Sciences
degois.publication.volume169
dspace.entity.typePublication
oaire.awardNumber205079
oaire.awardTitleUnlocking Animal Food Quality Research Potential in Baltic Region by Developing Scientific and Technical Capacities of the Research Institute “Sigra”
oaire.awardURIinfo:eu-repo/grantAgreement/EC/FP7/205079/EU
oaire.fundingStreamFP7
project.funder.identifierhttp://doi.org/10.13039/501100008530
project.funder.nameEuropean Commission
rcaap.rightsopenAccess
relation.isProjectOfPublication68fecc7d-49c2-4e11-acdd-5847c1229736
relation.isProjectOfPublication.latestForDiscovery68fecc7d-49c2-4e11-acdd-5847c1229736

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