Neutrophil elastase inhibitor purification strategy from cowpea seeds

Autor(es)

Ferreira, Graziele Cristina ^[1] ; Alves Duran, Adriana Feliciano ^[1] ; Santos da Silva, Flavia Ribeiro ^[1] ; Bomediano, Livia de Moraes ^[1] ; Machado, Gabriel Capella ^[1] ; Sasaki, Sergio Daishi ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed ABC, Ctr Ciencias Nat & Humanas, Sao Bernardo Do Campo, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: PLoS One; v. 14, n. 10 OCT 10 2019.
• Citações Web of Science: 0

Resumo

Serine proteases and its inhibitors are involved in physiological process and its deregulation lead to various diseases like Chronic Obstructive Pulmonary Disease (COPD), pulmonary emphysema, skin diseases, atherosclerosis, coagulation diseases, cancer, inflammatory diseases, neuronal disorders and other diseases. Serine protease inhibitors have been described in many species, as well as in plants, including cowpea beans (Vigna unguiculata (L.) Walp). Here, we purified and characterized a protease inhibitor, named VuEI (Vigna unguiculata elastase inhibitor), from Vigna unguiculata, with inhibitory activity against HNE (human neutrophil elastase) and chymotrypsin but has no inhibitory activity against trypsin and thrombin. VuEI was obtained by alkaline protein extraction followed by three different chromatographic steps in sequence. First, an ion exchange chromatography using Hitrap Q column was employed, followed by two reversed-phase chromatography using Source15RPC and ACE18 columns. The molecular mass of VuEI was estimated in 10.99 kDa by MALDI-TOF mass spectrometry. The dissociation constant (Ki) to HNE was 9 pM. These data indicate that VuEI is a potent inhibitor of human neutrophil elastase, besides to inhibit chymotrypsin. (AU)

• Processo FAPESP: 11/07001-7 - Estudos da atividade de inibidores de serinoproteases originários do carrapato Rhipicephalus Boophilus microplus em modelo de enfisema pulmonar em camundongos e caracterização molecular das atividades de serinoproteases presentes no modelo
• Beneficiário: Sergio Daishi Sasaki
• Modalidade de apoio: Auxílio à Pesquisa - Regular