Neutrophil elastase inhibitor purification strategy from cowpea seeds

Autoři: Graziele Cristina Ferreira aff001;  Adriana Feliciano Alves Duran aff001;  Flavia Ribeiro Santos da Silva aff001;  Livia de Moraes Bomediano aff001;  Gabriel Capella Machado aff001;  Sergio Daishi Sasaki aff001
Působiště autorů: Centro de Ciências Naturais e Humanas, Universidade Federal do ABC, São Bernardo do Campo, São Paulo, Brazil aff001
Vyšlo v časopise: PLoS ONE 14(10)
Kategorie: Research Article


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.

Klíčová slova:

Enzyme inhibitors – Chymotrypsin – Inflammatory diseases – Neutrophils – Proteases – Protein extraction – Serine proteases – Ion exchange chromatography


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2019 Číslo 10
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