Neutrophil elastase inhibitor purification strategy from cowpea seeds.
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...
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2019-01-01
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Series: | PLoS ONE |
Online Access: | https://doi.org/10.1371/journal.pone.0223713 |
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doaj-49fc921d334149c79661bc4f077d37682021-03-03T21:06:20ZengPublic Library of Science (PLoS)PLoS ONE1932-62032019-01-011410e022371310.1371/journal.pone.0223713Neutrophil elastase inhibitor purification strategy from cowpea seeds.Graziele Cristina FerreiraAdriana Feliciano Alves DuranFlavia Ribeiro Santos da SilvaLivia de Moraes BomedianoGabriel Capella MachadoSergio Daishi SasakiSerine 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.https://doi.org/10.1371/journal.pone.0223713 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Graziele Cristina Ferreira Adriana Feliciano Alves Duran Flavia Ribeiro Santos da Silva Livia de Moraes Bomediano Gabriel Capella Machado Sergio Daishi Sasaki |
spellingShingle |
Graziele Cristina Ferreira Adriana Feliciano Alves Duran Flavia Ribeiro Santos da Silva Livia de Moraes Bomediano Gabriel Capella Machado Sergio Daishi Sasaki Neutrophil elastase inhibitor purification strategy from cowpea seeds. PLoS ONE |
author_facet |
Graziele Cristina Ferreira Adriana Feliciano Alves Duran Flavia Ribeiro Santos da Silva Livia de Moraes Bomediano Gabriel Capella Machado Sergio Daishi Sasaki |
author_sort |
Graziele Cristina Ferreira |
title |
Neutrophil elastase inhibitor purification strategy from cowpea seeds. |
title_short |
Neutrophil elastase inhibitor purification strategy from cowpea seeds. |
title_full |
Neutrophil elastase inhibitor purification strategy from cowpea seeds. |
title_fullStr |
Neutrophil elastase inhibitor purification strategy from cowpea seeds. |
title_full_unstemmed |
Neutrophil elastase inhibitor purification strategy from cowpea seeds. |
title_sort |
neutrophil elastase inhibitor purification strategy from cowpea seeds. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2019-01-01 |
description |
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. |
url |
https://doi.org/10.1371/journal.pone.0223713 |
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