We have purified to homogeneity two forms of a new serine protease inhibitor specific for elastase/chymotrypsin from the ovary gland of the desert locust Schistocerca gregaria. This protein, greglin, has 83 amino acid residues and bears putative phosphorylation sites. Amino acid sequence alignments revealed no homology with pacifastin insect inhibitors and only a distant relationship with Kazal-type inhibitors. This was confirmed by computer-based structural studies. The most closely related homologue is a putative gene product from Ciona intestinalis with which it shares 38% sequence homology. Greglin is a fast-acting and tight binding inhibitor of human neutrophil elastase (kass=1.2×107 M−1·s−1, Ki=3.6 nM) and subtilisin. It also binds neutrophil cathepsin G, pancreatic elastase and chymotrypsin with a lower affinity (26 nM≤Ki≤153 nM), but does not inhibit neutrophil protease 3 or pancreatic trypsin. The capacity of greglin to inhibit neutrophil elastase was not significantly affected by exposure to acetonitrile, high temperature (90 °C), low or high pH (2.5–11.0), N-chlorosuccinimide-mediated oxidation or the proteolytic enzymes trypsin, papain and pseudolysin from Pseudomonas aeruginosa. Greglin efficiently inhibits the neutrophil elastase activity of sputum supernatants from cystic fibrosis patients. Its biological function in the locust ovary gland is currently unknown, but its physicochemical properties suggest that it can be used as a template to design a new generation of highly resistant elastase inhibitors for treating inflammatory diseases.
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December 2006
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Research Article|
November 28 2006
A novel locust (Schistocerca gregaria) serine protease inhibitor with a high affinity for neutrophil elastase
Michèle Brillard-Bourdet;
Michèle Brillard-Bourdet
*INSERM U618, F-37000 Tours, France
†Université François Rabelais, F-37000 Tours, France
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Ahmed Hamdaoui;
Ahmed Hamdaoui
‡Université Cadi Ayyad, Faculté des Sciences Semlalia, Marrakech, Morocco
§INSERM U392, Université Louis Pasteur, Strasbourg I, F-67401 Illkirch, France
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Eric Hajjar;
Eric Hajjar
∥Computational Biology Unit, BCCS, University of Bergen, N-5008 Bergen, Norway
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Christian Boudier;
Christian Boudier
¶CNRS UMR 7175, Université Louis Pasteur, Strasbourg I, F-67401 Illkirch, France
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Nathalie Reuter;
Nathalie Reuter
∥Computational Biology Unit, BCCS, University of Bergen, N-5008 Bergen, Norway
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Laurence Ehret-Sabatier;
Laurence Ehret-Sabatier
**CNRS UMR 7512, Ecole de Chimie des Polymères et Matériaux, F-67087 Strasbourg, France
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Joseph G. Bieth;
Joseph G. Bieth
§INSERM U392, Université Louis Pasteur, Strasbourg I, F-67401 Illkirch, France
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Francis Gauthier
Francis Gauthier
1
*INSERM U618, F-37000 Tours, France
†Université François Rabelais, F-37000 Tours, France
1To whom correspondence should be addressed, at INSERM U618 ‘Protéases et Vectorisation Pulmonaires’, Université François Rabelais, 10 Bd Tonnellé, 37032 Tours Cedex, France (email gauthier@univ-tours.fr).
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Publisher: Portland Press Ltd
Received:
March 22 2006
Revision Received:
July 10 2006
Accepted:
July 13 2006
Accepted Manuscript online:
July 13 2006
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2006
Biochem J (2006) 400 (3): 467–476.
Article history
Received:
March 22 2006
Revision Received:
July 10 2006
Accepted:
July 13 2006
Accepted Manuscript online:
July 13 2006
Citation
Michèle Brillard-Bourdet, Ahmed Hamdaoui, Eric Hajjar, Christian Boudier, Nathalie Reuter, Laurence Ehret-Sabatier, Joseph G. Bieth, Francis Gauthier; A novel locust (Schistocerca gregaria) serine protease inhibitor with a high affinity for neutrophil elastase. Biochem J 15 December 2006; 400 (3): 467–476. doi: https://doi.org/10.1042/BJ20060437
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