A novel cysteine proteinase inhibitor from seeds of Enterolobium contortisiliquum and its effect on Callosobruchus maculatus larvae

Autor(es)

Nunes, Natalia N. S. ^[1] ; Ferreira, Rodrigo S. ^[1] ; de Sa, Leonardo F. R. ^[2] ; de Oliveira, Antonia Elenir A. ^[2] ; Oliva, V, Maria Luiza

Afiliação do(s) autor(es)

^[1] V, Univ Fed Sao Paulo UNIFESP, Biochem Dept, BR-04044020 Sao Paulo, SP - Brazil

^[2] Univ Estadual Norte Fluminense Darcy Ribeiro UENF, Biosci & Biotechnol Ctr, Chem Lab Chem & Funct Prot & Peptides, CBB, Campos Dos Goytacazes, RJ - Brazil

• Tipo de documento: Artigo Científico
• Fonte: BIOCHEMISTRY AND BIOPHYSICS REPORTS; v. 25, MAR 2021.
• Citações Web of Science: 0

Resumo

This study focused on the characterization of a novel cysteine proteinase inhibitor from Enterolobium contortisiliquum seeds targeting the inhibition of the growth of Callosobruchus maculatus larvae, an important cosmopolitan pest of the cowpea Vigna unguiculata during storage. The inhibitor was isolated by ion-exchange besides of size exclusion chromatography. EcCI molecular mass is 19,757 Da, composed of two polypeptide chains. It strongly inhibits papain (Ki(app) 0.036 nM) and proteinases from the midguts of C. maculatus (80 mu g mL(-1), 60% inhibition). The inhibitory activity is reduced by 40% after a heat treatment at 100 degrees C for 2 h. The protein displayed noxious activity at 0.5% and 1% (w/w) when incorporated in artificial seeds, reducing larval mass in 87% and 92%, respectively. Treatment of C. maculatus larvae with conjugated EcCI-FIT and subsequent biodistribution resulted in high fluorescence intensity in midguts and markedly low intensity in malpighian tubules and fat body. Small amounts of labeled proteins were detected in larvae feces. The detection of high fluorescence in larvae midguts and low fluorescence in their feces indicate the retention of the FITC conjugated EcCI inhibitor in larvae midguts. These results demonstrate the potential of the natural protein from E. contortisiliquum to inhibit the development of C. maculatus. (AU)

• Processo FAPESP: 17/07972-9 - Desenvolvimento de compostos para profilaxia e tratamento de doenças cardiovasculares
• Beneficiário: Maria Luiza Vilela Oliva
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 17/06630-7 - Fragmentos derivados de proteínas com seletividade para inibição de enzimas de mamíferos e micro-organismos e seu papel como agente anti-inflamatório, antimicrobiano, antitrombótico e antitumoral: mecanismo de ação
• Beneficiário: Maria Luiza Vilela Oliva
• Modalidade de apoio: Auxílio à Pesquisa - Temático