scholarly journals A Versatile and Robust Serine Protease Inhibitor Scaffold from Actinia tenebrosa

Marine Drugs ◽  
2019 ◽  
Vol 17 (12) ◽  
pp. 701 ◽  
Author(s):  
Xingchen Chen ◽  
Darren Leahy ◽  
Jessica Van Haeften ◽  
Perry Hartfield ◽  
Peter J. Prentis ◽  
...  
Keyword(s):  
Serine Protease ◽  
Protease Inhibitors ◽  
Serine Proteases ◽  
Inhibitor Design ◽  
Consensus Analysis ◽  
Fine Grained ◽  
Human Sequence ◽  
Physiological Processes ◽  
Definition Of ◽  
Dynamics Simulations

Serine proteases play pivotal roles in normal physiology and a spectrum of patho-physiological processes. Accordingly, there is considerable interest in the discovery and design of potent serine protease inhibitors for therapeutic applications. This led to concerted efforts to discover versatile and robust molecular scaffolds for inhibitor design. This investigation is a bioprospecting study that aims to isolate and identify protease inhibitors from the cnidarian Actinia tenebrosa. The study isolated two Kunitz-type protease inhibitors with very similar sequences but quite divergent inhibitory potencies when assayed against bovine trypsin, chymostrypsin, and a selection of human sequence-related peptidases. Homology modeling and molecular dynamics simulations of these inhibitors in complex with their targets were carried out and, collectively, these methodologies enabled the definition of a versatile scaffold for inhibitor design. Thermal denaturation studies showed that the inhibitors were remarkably robust. To gain a fine-grained map of the residues responsible for this stability, we conducted in silico alanine scanning and quantified individual residue contributions to the inhibitor’s stability. Sequences of these inhibitors were then used to search for Kunitz homologs in an A. tenebrosa transcriptome library, resulting in the discovery of a further 14 related sequences. Consensus analysis of these variants identified a rich molecular diversity of Kunitz domains and expanded the palette of potential residue substitutions for rational inhibitor design using this domain.

Regulation of kallikrein-related peptidases in the skin – from physiology to diseases to therapeutic options

2013 ◽  
Vol 110 (09) ◽  
pp. 442-449 ◽  
Author(s):  
Jan Fischer ◽  
Ulf Meyer-Hoffert
Keyword(s):  
Serine Protease ◽  
Protease Inhibitors ◽  
Therapeutic Intervention ◽  
Serine Proteases ◽  
Current Knowledge ◽  
Serine Protease Inhibitors ◽  
Specific Expression ◽  
Physiological Processes ◽  
Promising Target ◽  
Protease Activation

SummaryKallikrein-related peptidases (KLKs) constitute a family of 15 highly conserved serine proteases, which show a tissue-specific expression profile. This made them valuable tumour expression markers. It became evident that KLKs are involved in many physiological processes like semen liquefaction and skin desquamation. More recently, we have learnt that they are involved in many pathophysiological conditions and diseases making them promising target of therapeutic intervention. Therefore, regulation of KLKs raised the interest of numerous reports. Herein, we summarise the current knowledge on KLKs regulation with an emphasis on skin-relevant KLKs regulation processes. Regulation of KLKs takes place on the level of transcription, on protease activation and on protease inactivation. A variety of protease inhibitors has been described to interact with KLKs including the irreversible serine protease inhibitors (SERPINs) and the reversible serine protease inhibitors of Kazal-type (SPINKs). In an attempt to integrate current knowledge, we propose that KLK regulation has credentials as targets for therapeutic intervention.

scholarly journals Serine proteases and serine protease inhibitors in testicular physiology: the plasminogen activation system

Reproduction ◽  
2007 ◽  
Vol 134 (6) ◽  
pp. 721-729 ◽  
Author(s):  
Brigitte Le Magueresse-Battistoni
Keyword(s):  
Serine Protease ◽  
Protease Inhibitors ◽  
Expression Pattern ◽  
Serine Proteases ◽  
Current Knowledge ◽  
Adult Life ◽  
Plasminogen Activation ◽  
Serine Protease Inhibitors ◽  
Plasminogen Activation System ◽  
Activation System

The testis is an organ in which a series of radical remodeling events occurs during development and in adult life. These events likely rely on a sophisticated network of proteases and complementary inhibitors, including the plasminogen activation system. This review summarizes our current knowledge on the testicular occurrence and expression pattern of members of the plasminogen activation system. The various predicted functions for these molecules in the establishment and maintenance of the testicular architecture and in the process of spermatogenesis are presented.

scholarly journals Therapeutic targeting of coronavirus spike glycoprotein priming

2020 ◽  
Author(s):  
Maurizio Pellecchia ◽  
Elisa Barile ◽  
Carlo Baggio ◽  
Luca Gambini ◽  
Sergey A. Shiryaev ◽  
...  
Keyword(s):  
Serine Protease ◽  
Protease Inhibitors ◽  
Serine Proteases ◽  
Viral Proteins ◽  
Exact Nature ◽  
Cleavage Sites ◽  
Serine Protease Inhibitors ◽  
Spike Glycoprotein ◽  
Highly Pathogenic

Abstract Processing of certain viral proteins and bacterial toxins by host serine proteases is a frequent and critical step in virulence. The coronavirus spike glycoprotein contains three (S1, S2, and S2’) cleavage sites that are processed by human host proteases. The exact nature of these cleavage sites, and their respective processing proteases, can determine whether the virus can cross species, and the level of pathogenicity. Recent comparisons of the genomes of the highly pathogenic SARS-CoV2 and MERS-CoV, with less pathogenic strains (e.g., Bat-RaTG13, the bat homologue of SARS-CoV2) identified possible mutations in the receptor binding domain and in the S1 and S2’ cleavage sites of their spike glycoprotein. However there remains some confusion on the relative roles of the possible serine-proteases involved for priming. Using anthrax toxin as a model system, we show that in vivo inhibition of priming by pan-active serine protease inhibitors can be effective at suppressing virulence. Hence, our studies should encourage further efforts in developing either pan-serine protease inhibitors or inhibitor cocktails to target SARS-CoV2 and potentially ward off future pandemics that could develop because of the additional mutations in the S-protein priming sequence in coronaviruses.

scholarly journals Serine protease inhibitors and human wellbeing interplay: new insights for old friends

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e7224 ◽  
Author(s):  
Héla Mkaouar ◽  
Nizar Akermi ◽  
Aicha Kriaa ◽  
Anne-Laure Abraham ◽  
Amin Jablaoui ◽  
...  
Keyword(s):  
Serine Protease ◽  
Protease Inhibitors ◽  
Structural Data ◽  
Ecological Niches ◽  
Serine Protease Inhibitors ◽  
New Therapeutic Approaches ◽  
Physiological Processes ◽  
Health And Disease ◽  
And Function

Serine Protease Inhibitors (Serpins) control tightly regulated physiological processes and their dysfunction is associated to various diseases. Thus, increasing interest is given to these proteins as new therapeutic targets. Several studies provided functional and structural data about human serpins. By comparison, only little knowledge regarding bacterial serpins exists. Through the emergence of metagenomic studies, many bacterial serpins were identified from numerous ecological niches including the human gut microbiota. The origin, distribution and function of these proteins remain to be established. In this report, we shed light on the key role of human and bacterial serpins in health and disease. Moreover, we analyze their function, phylogeny and ecological distribution. This review highlights the potential use of bacterial serpins to set out new therapeutic approaches.

Alpha-1 Antitrypsin Is a Potent Inhibitor of Cytokine-Induced Hematopoietic Stem Cell Mobilization.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1185-1185
Author(s):  
Melissa van Pel ◽  
Ronald van Os ◽  
Gerjo A. Velders ◽  
Henny Hagoort ◽  
Ivan J. Lindley ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Serine Protease ◽  
Protease Inhibitors ◽  
Serine Proteases ◽  
Low Dose ◽  
Serine Protease Inhibitors ◽  
Hematopoietic Stem ◽  
Substrate Conversion

Abstract Previously, we have shown that IL-8 and G-CSF-induced hematopoietic stem cell (HSC) mobilization is inhibited in mice that underwent low dose (0.5 Gy) total body irradiation (TBI), whereas the number of progenitor cells in the bone marrow remained similar in all groups. The mechanism underlying this inhibition remains unknown. Since the release of granular proteases by neutrophils is well known to play a role in HSC mobilization, we also considered a possible role for serine protease inhibitors in the induction of HSC mobilization. Serine proteases, such as elastase and cathepsin G, are irreversibly inhibited by serine protease inhibitors including alpha-1 antitrypsin (alpha-1 AT) and alpha2-macroglobulin. In-vitro tests revealed that addition of bone marrow extracellular extracts, that were obtained from murine femurs 24 hours following low dose (0.5 Gy) TBI, inhibited the activity of exogenous elastase in a chromogenic substrate conversion assay up to 78.1 % compared to extracts obtained from sham irradiated controls (p<0.05). Since elastase inhibition by alpha2-macroglobulin cannot be detected in a chromogenic substrate conversion assay, alpha-1 AT was considered as the primary candidate serine protease inhibitor to inhibit elastase activity in our in-vitro system. Quantitative PCR of total bone marrow cells revealed that alpha-1 AT mRNA was 20-fold increased relative to the housekeeping gene ß-actin and 7-fold relative to the housekeeping genes HPRT and GAPDH at 24 hours following low dose (0.5 Gy) TBI. In addition, Western blot analysis indicated that alpha-1 AT protein concentrations were significantly (p<0.01) increased in bone marrow extracellular extracts derived from low dose (0.5 Gy) irradiated mice, compared to extracts obtained from sham-irradiated controls (5.1 ± 0.6 scanning units [SU] vs. 3.9 ± 0.7 SU for 0.5 Gy;n=8 vs. 0 Gy; n=6 respectively). To further substantiate a possible in-vivo role of alpha-1 AT in the inhibition of HSC mobilization, we administered alpha-1 AT (300 μg/mouse i.p.) at 2 hours and at 5 minutes prior to IL-8 injection (30 μg/mouse i.p.). Administration of alpha-1 AT prior to IL-8 injection completely (p<0.05) inhibited IL-8-induced HSC mobilization (472.9 ± 289.5 CFU-GM per ml blood for IL-8; n=5 vs. 44.8 ± 35.5 CFU-GM per ml blood for alpha-1 AT/IL-8; n =11). These results indicate that 1) alpha-1 AT is a potent inhibitor of IL-8-induced HSC mobilization and 2) in-vivo induced alpha-1 AT contributes to the inhibition of HSC mobilization after low-dose (0.5 Gy) TBI. We hypothesize that a critical balance between serine proteases and serine protease inhibitors plays an important role in cytokine-induced HSC mobilization.

scholarly journals Activity of serine proteases from Fasciola hepatica eggs in relation to pH and temperature

2020 ◽  
Vol 23 (3) ◽  
pp. 350-358
Author(s):  
L. Kianifard ◽  
M. Yakhchali ◽  
M. Imani
Keyword(s):  
Serine Protease ◽  
Protease Inhibitors ◽  
Proteolytic Activity ◽  
Serine Proteases ◽  
Fasciola Hepatica ◽  
Ethylenediaminetetraacetic Acid ◽  
Optimum Temperature ◽  
Egg Hatching ◽  
Protein Levels ◽  
Ph Values

This study was conducted to analyse the serine protease of Fasciola hepatica eggs by specific substrates and inhibitors, and investigation of the effects of pH and temperature on proteases’ activity and stability. Adult worms were isolated from infected livers. After homogenisation, their protein levels were measured with the Bradford method. Total proteolytic activity of the Fasciola hepatica extract was evaluated with azocasein substrate at pH values from 2 to 12. N-benzoyl–arginine–p-nitroanilide (BApNA) trypsin and N-succinyl-alanine-alanine-prolin-phenylalanine-p-nitroanilide (SAAPFpNA) chymotrypsin substrates were used to measure specific protease activities. The effect of protease inhibitors phenylmethane sulfonyl fluoride (PMSF), pepsin, and ethylenediaminetetraacetic acid (EDTA) on these enzymes was evaluated. Estimation of optimum temperature and pH was performed in the temperature range of 10–90 °C and pH values from 2–12. The optimum pH activities for trypsin and chymotrypsin were at alkaline pH and for total proteolytic activity at acidic pH. The results using protease inhibitors showed that the eggs had serine protease activity. The optimum temperature activity of trypsin and chymotrypsin was 50 °C. These proteases were stable up to 40 °C. Due to the importance of pH and temperature in life cycle of Fasciola hepatica, these findings can be used for induction of some modifications in pH and preventing the activity of the enzyme for decrement of the efficacy of embryonic development and egg hatching of this zoonotic parasite.

scholarly journals Interdependence of kallikrein-related peptidases in proteolytic networks

2010 ◽  
Vol 391 (5) ◽  
pp. 581-587 ◽  
Author(s):  
Nathalie Beaufort ◽  
Karolina Plaza ◽  
Daniel Utzschneider ◽  
Amelie Schwarz ◽  
Julia M. Burkhart ◽  
...  
Keyword(s):  
Matrix Metalloproteinase ◽  
Serine Protease ◽  
Serine Proteases ◽  
Matrix Metalloproteinase 3 ◽  
Physiological Processes

Abstract Human kallikrein-related peptidases (KLKs) are 15 homologous serine proteases involved in several (patho)physiological processes, including cancer. Secreted as precursors, they are activated upon proteolytic release of a short pro-peptide. We searched for interconnection of KLKs within extracellular proteolytic networks leading to activation of protease zymogens and found that (i) pro-KLK activation by other KLKs is scarce, with the exception of pro-KLK11, which is efficiently activated by KLK4 and 5; (ii) pro-KLK4 is activated by matrix metalloproteinase 3; and (iii) trypsin-like KLKs efficiently activate the serine protease urokinase. Our observations provide new insights into the regulation of these important tumor-associated proteases.

scholarly journals Prospects for the design of new therapeutically significant protease inhibitors based on knottins and sunflower seed trypsin inhibitor (SFTI 1)

2016 ◽  
Vol 62 (4) ◽  
pp. 353-368 ◽  
Author(s):  
S.S. Kuznetsova ◽  
E.F. Kolesanova ◽  
A.V. Talanova ◽  
A.V. Veselovsky
Keyword(s):  
Serine Protease ◽  
Protease Inhibitors ◽  
Trypsin Inhibitor ◽  
Serine Proteases ◽  
Sunflower Seed ◽  
High Efficiency ◽  
Structural Characteristics ◽  
Research Field ◽  
Amino Acid Sequences ◽  
Plant Seed

Plant seed knottins, mainly from the Cucurbitacea family, and sunflower seed trypsin inhibitor (SFTI 1) are the most low-molecular canonical peptide inhibitors of serine proteases. High efficiency of inhibition of various serine proteases, structure rigidity together with the possibility of limited variations of amino acid sequences, high chemical stability, lack of toxic properties, opportunity of production by either chemical synthesis or use of heterologous expression systems make these inhibitors attractive templates for design of new compounds for regulation of therapeutically significant serine protease activities. Hence the design of such compounds represents a prospective research field. The review considers structural characteristics of these inhibitors, their properties, methods of preparation and design of new analogs. Examples of successful employment of natural serine protease inhibitors belonging to knottin family and SFTI 1 as templates for the design of highly specific inhibitors of certain proteases are given.

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