Increased Stability of Oligopeptidases Immobilized on Gold Nanoparticles

Autor(es)

Icimoto, Marcelo Yudi ^[1] ; Brito, Adrianne Marlise Mendes ^{[1, 2]} ; Ramos, Marcos Paulo Cyrillo ^[1] ; Oliveira, Vitor ^[1] ; Nantes-Cardoso, Iseli Lourenco ^[2]

Afiliação do(s) autor(es)

^[1] Univ Fed Sao Paulo, Dept Biofis, BR-04024002 Sao Paulo - Brazil

^[2] Univ Fed ABC, Ctr Ciencias Naturais & Humanas, BR-09210580 Santo Andre, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: CATALYSTS; v. 10, n. 1 JAN 2020.
• Citações Web of Science: 1

Resumo

The metallopeptidases thimet oligopeptidase (THOP, EC 3.4.24.25) and neurolysin (NEL, EC 3.4.24.26) are enzymes that belong to the zinc endopeptidase M13 family. Numerous studies suggest that these peptidases participate in the processing of bioactive peptides such as angiotensins and bradykinin. Efforts have been conducted to develop biotechnological tools to make possible the use of both proteases to regulate blood pressure in mice, mainly limited by the low plasmatic stability of the enzymes. In the present study, it was investigated the use of nanotechnology as an efficient strategy for to circumvent the low stability of the proteases. Recombinant THOP and NEL were immobilized in gold nanoparticles (GNPs) synthesized in situ using HEPES and the enzymes as reducing and stabilizing agents. The formation of rTHOP-GNP and rNEL-GNP was characterized by the surface plasmon resonance band, zeta potential and atomic force microscopy. The gain of structural stability and activity of rTHOP and rNEL immobilized on GNPs was demonstrated by assays using fluorogenic substrates. The enzymes were also efficiently immobilized on GNPs fabricated with sodium borohydride. The efficient immobilization of the oligopeptidases in gold nanoparticles with gain of stability may facilitate the use of the enzymes in therapies related to pressure regulation and stroke, and as a tool for studying the physiological and pathological roles of both proteases. (AU)

• Processo FAPESP: 19/01487-7 - Nanopartículas de ouro associadas à cisteíno-proteases derivadas de Bromelaína: caracterização estrutural e ação colagenolítica
• Beneficiário: Marcelo Yudi Icimoto
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 17/02317-2 - Interfaces em materiais: propriedades eletrônicas, magnéticas, estruturais e de transporte
• Beneficiário: Adalberto Fazzio
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 18/09158-0 - Prolil oligopeptidase: estrutura-atividade e secreção
• Beneficiário: Vitor Marcelo Silveira Bueno Brandão de Oliveira
• Modalidade de apoio: Auxílio à Pesquisa - Regular