Artigo De: orcid, cienciavitae

Recognition of nucleolin through interaction with RNA G-quadruplex

Biochemical Pharmacology

Santos, Tiago; Miranda, André; Cruz, Carla2021Elsevier

Informações chave

Autores:

Santos, Tiago; Miranda, André; Campello, Maria P.C. (Maria Paula Cordeiro Crespo Cabral Campello Aboim de Barros); Paulo, António (António Manuel Rocha Paulo); Salgado, Gilmar; Cabrita, Eurico J.; Cruz, Carla

Publicado em

Julho 2021

Resumo

The development of novel biomarkers for early-stage diagnosis of prostate cancer (PCa) has attracted the attention of researchers in the last decade. Nucleolin (NCL) has emerged as a possible biomarker of PCa due to its high expression levels in the surface of PCa cells and affinity towards parallel G4s since it contains four RNA-binding domains (RBDs). Herein, we developed a novel strategy based on a microfluidic platform for the detection of NCL in biological samples, such as human plasma. The RNA G4 (rG4) sequence found in human precursor microRNA 92b (pre-miR-92b) was used as a molecular recognition probe since it forms a single dominant parallel rG4 conformation in the presence of 0.1 mM K+ as confirmed by NMR spectroscopy. The additional stability of the rG4 structure was provided by the acridine orange derivative ligand C8, which stabilizes the pre-miR-92b rG4 structure, as denoted by an increase in more than 30 °C of its melting temperature. FRET-melting assay revealed a remarkable synergistic effect of NCL RBD1,2 and C8 on the stabilization of the pre-miR-92b rG4. The binding of pre-miR-92b to NCL RBD1,2 was determined by in silico studies, which revealed a binding pocket formed by a 12-residue linker between RBD1 and RBD2. Both, pre-miR-92b rG4 and pre-miR-92b rG4/C8 complex demonstrated high affinity towards NCL RBD1,2, as proved by fluorimetric titrations (KD range between 10-12 and 10-9 M). The stability and nuclease resistance of pre-miR-92b rG4 and pre-miR-92b rG4/C8 complex were evaluated as molecular recognition probes to capture and detect NCL. Finally, the microfluidic platform detects NCL in complex biological samples, such as human plasma. Overall, this work demonstrates the usefulness of the microfluidic platform based on the pre-miR-92b to detect NCL and the possibility to be used as a valuable biomedical tool in PCa diagnosis.

Detalhes da publicação

Autores da comunidade :

Versão da publicação

AO - Versão original do autor

Editora

Elsevier

Ligação para a versão da editora

https://www.sciencedirect.com/journal/biochemical-pharmacology

Título do contentor da publicação

Biochemical Pharmacology

Primeira página ou número de artigo

114208

Volume

189

ISSN

0006-2952

WoS (Web of Science)

WOS:000662577800008

Domínio Científico (FOS)

chemical-sciences - Química

Palavras-chave

  • Nucleolin
  • RNA G-quadruplex
  • Acridine ligands
  • Microfluidics and Prostate cancer

Idioma da publicação (código ISO)

eng - Inglês

Identificador alternativo (URI)

http://dx.doi.org/10.1016/j.bcp.2020.114208

Acesso à publicação:

Acesso apenas a metadados