A novel locust (Schistocerca gregaria) serine protease inhibitor with a high affinity for neutrophil elastase.
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 (k(ass)=1.2x10(7) M(-1) x s(-1), K(i)=3.6 nM) and subtilisin. It also binds neutrophil cathepsin G, pancreatic elastase and chymotrypsin with a lower affinity (26 nM< or =K(i)< or =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 degrees 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.
Brillard-Bourdet
Michèle
M
INSERM U618, F-37000 Tours, France.
Hamdaoui
Ahmed
A
Hajjar
Eric
E
Boudier
Christian
C
Reuter
Nathalie
N
Ehret-Sabatier
Laurence
L
Bieth
Joseph G
JG
Gauthier
Francis
F
D000595:Amino Acid Sequence; D000818:Animals; D005260:Female; D006110:Grasshoppers / Q000737:chemistry*; D019272:Leukocyte Elastase / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D008958:Models, Molecular; D010053:Ovary / Q000378:metabolism; D011487:Protein Conformation; D020033:Protein Isoforms; D015842:Serine Proteinase Inhibitors / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*
D020033:Protein Isoforms; D015842:Serine Proteinase Inhibitors; D019272:Leukocyte Elastase