scholarly journals A novel locust (Schistocerca gregaria) serine protease inhibitor with a high affinity for neutrophil elastase

2006 ◽  
Vol 400 (3) ◽  
pp. 467-476 ◽  
Author(s):  
Michèle Brillard-Bourdet ◽  
Ahmed Hamdaoui ◽  
Eric Hajjar ◽  
Christian Boudier ◽  
Nathalie Reuter ◽  
...  
Keyword(s):  
Amino Acid ◽  
Protease Inhibitor ◽  
Serine Protease ◽  
Neutrophil Elastase ◽  
Schistocerca Gregaria ◽  
Serine Protease Inhibitor ◽  
Cathepsin G ◽  
Human Neutrophil Elastase ◽  
Sequence Alignments ◽  
Putative Gene

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.

scholarly journals “Sleeping Beauty” Phenomenon: SuFEx-Enabled Discovery of Selective Covalent Inhibitors of Human Neutrophil Elastase

2019 ◽  
Author(s):  
Qinheng Zheng ◽  
Jordan L. Woehl ◽  
Seiya Kitamura ◽  
Diogo Santos-Martins ◽  
Christopher J. Smedley ◽  
...  
Keyword(s):  
Serine Protease ◽  
Neutrophil Elastase ◽  
Human Neutrophil ◽  
Sleeping Beauty ◽  
Cathepsin G ◽  
Human Neutrophil Elastase ◽  
Biological Assays ◽  
Covalent Inhibitors ◽  
Sulfur Fluoride ◽  
New Generation

<div> <div> <div> <p>Sulfur-Fluoride Exchange (SuFEx) has emerged as the new generation of click chemistry. We report here a SuFEx-enabled approach exploiting the “sleeping beauty” phenomenon of sulfur fluoride compounds in the context of the serendipitous discovery of selective covalent human neutrophil elastase (hNE) inhibitors. Evaluation of an ever-growing collection of SuFExable compounds toward various biological assays unexpectedly yielded a selective and covalent hNE inhibitor, benzene-1,2-disulfonyl fluoride. Derivatization of the initial hit led to a better agent, 2- triflyl benzenesulfonyl fluoride, itself made through a SuFEx trifluoromethylation process, with IC50 = 1.1 μM and ~200-fold selectivity over the homologous neutrophil serine protease, cathepsin G. The optimized probe only modified active hNE and not its denatured form, setting another example of the “sleeping beauty” phenomenon of sulfur fluoride capturing agents for the discovery of covalent medicines. </p> </div> </div> </div>

scholarly journals Structural modeling and thermostability of a serine protease inhibitor belonging to the Kunitz family from the tick Rhipicephalus microplus

2020 ◽  
Author(s):  
Lívia de Moraes Bomediano Camillo ◽  
Graziele Cristina Ferreira ◽  
Adriana Feliciano Alves Duran ◽  
Flavia Ribeiro Santos da Silva ◽  
Wanius Garcia ◽  
...  
Keyword(s):  
Protease Inhibitor ◽  
Serine Protease ◽  
Disulfide Bonds ◽  
Tertiary Structure ◽  
Dissociation Constants ◽  
Serine Protease Inhibitor ◽  
Rhipicephalus Microplus ◽  
Human Neutrophil Elastase ◽  
Structure Stabilization

ABSTRACTrBmTI-A is a recombinant serine protease inhibitor that belongs to the Kunitz-BPTI family and that was cloned from Rhipicephalus microplus tick. rBmTI-A has inhibitory activities on bovine trypsin, human plasma kallikrein, human neutrophil elastase and plasmin with dissociation constants in nM range. It is characterized by two inhibitory domains and each domain presents six cysteines that form three disulfide bonds, which contribute to the high stability of its structure. Previous studies suggest that serine protease inhibitor rBmTI-A has a protective potential against pulmonary emphysema in mice and anti-inflammatory potential, besides rBmTI-A presented a potent inhibitory activity against in vitro vessel formation. In this study, the tertiary structure of BmTI-A was modeled based on the structure of its Sabellastarte magnifica homologue. The structure stabilization was evaluated by molecular dynamics analysis. Circular dichroism data corroborated the secondary structure found by the homology modeling. Thermostability analysis confirmed the thermostability and the relation between the effects of the temperature in the inhibitor activity. The loss of activity observed was gradual, and, after 60 minutes of incubation at 90°C the inhibitor lost it completely.

scholarly journals SerpinB1 is critical for neutrophil survival through cell-autonomous inhibition of cathepsin G

Blood ◽  
2013 ◽  
Vol 121 (19) ◽  
pp. 3900-3907 ◽  
Author(s):  
Mathias Baumann ◽  
Christine T. N. Pham ◽  
Charaf Benarafa
Keyword(s):  
Protease Inhibitor ◽  
Serine Protease ◽  
Serine Protease Inhibitor ◽  
Cathepsin G ◽  
Key Points ◽  
Azurophil Granule ◽  
Neutrophil Survival

Key Points Serine protease inhibitor serpinB1 protects neutrophils by inhibition of their own azurophil granule protease cathepsin G. Granule permeabilization in neutrophils leads to cathepsin G–mediated death upstream and independent of apoptotic caspases.

First Archaeal PEPB-Serine Protease Inhibitor fromSulfolobus solfataricuswith Noncanonical Amino Acid Sequence in the Reactive-Site Loop

2009 ◽  
Vol 8 (1) ◽  
pp. 327-334 ◽  
Author(s):  
Gianna Palmieri ◽  
Giuliana Catara ◽  
Michele Saviano ◽  
Emma Langella ◽  
Marta Gogliettino ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Protease Inhibitor ◽  
Serine Protease ◽  
Serine Protease Inhibitor ◽  
Reactive Site ◽  
Noncanonical Amino Acid

scholarly journals SuFEx-enabled, agnostic discovery of covalent inhibitors of human neutrophil elastase

2019 ◽  
Vol 116 (38) ◽  
pp. 18808-18814 ◽  
Author(s):  
Qinheng Zheng ◽  
Jordan L. Woehl ◽  
Seiya Kitamura ◽  
Diogo Santos-Martins ◽  
Christopher J. Smedley ◽  
...  
Keyword(s):  
Click Chemistry ◽  
Serine Protease ◽  
Neutrophil Elastase ◽  
Human Neutrophil ◽  
Cathepsin G ◽  
Human Neutrophil Elastase ◽  
Biological Assays ◽  
Covalent Inhibitors ◽  
Sulfur Fluoride ◽  
New Generation

Sulfur fluoride exchange (SuFEx) has emerged as the new generation of click chemistry. We report here a SuFEx-enabled, agnostic approach for the discovery and optimization of covalent inhibitors of human neutrophil elastase (hNE). Evaluation of our ever-growing collection of SuFExable compounds toward various biological assays unexpectedly revealed a selective and covalent hNE inhibitor: benzene-1,2-disulfonyl fluoride. Synthetic derivatization of the initial hit led to a more potent agent, 2-(fluorosulfonyl)phenyl fluorosulfate with IC50 0.24 μM and greater than 833-fold selectivity over the homologous neutrophil serine protease, cathepsin G. The optimized, yet simple benzenoid probe only modified active hNE and not its denatured form.

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