scholarly journals Phenotypic Analysis of Mice Lacking the Tmprss2-Encoded Protease

2006 ◽  
Vol 26 (3) ◽  
pp. 965-975 ◽  
Author(s):  
Tom S. Kim ◽  
Cynthia Heinlein ◽  
Robert C. Hackman ◽  
Peter S. Nelson
Keyword(s):  
Serine Protease ◽  
Serine Proteases ◽  
Biological Activities ◽  
Type Ii ◽  
Phenotypic Analysis ◽  
Normal Prostate ◽  
Prostate Hyperplasia ◽  
Transmembrane Serine Protease

ABSTRACT Tmprss2 encodes an androgen-regulated type II transmembrane serine protease (TTSP) expressed highly in normal prostate epithelium and has been implicated in prostate carcinogenesis. Although in vitro studies suggest protease-activated receptor 2 may be a substrate for TMPRSS2, the in vivo biological activities of TMPRSS2 remain unknown. We generated Tmprss2 −/− mice by disrupting the serine protease domain through homologous recombination. Compared to wild-type littermates, Tmprss2 −/− mice developed normally, survived to adulthood with no differences in protein levels of prostatic secretions, and exhibited no discernible abnormalities in organ histology or function. Loss of TMPRSS2 serine protease activity did not influence fertility, reduce survival, result in prostate hyperplasia or carcinoma, or alter prostatic luminal epithelial cell regrowth following castration and androgen replacement. Lack of an observable phenotype in Tmprss2 −/− mice was not due to transcriptional compensation by closely related Tmprss2 homologs. We conclude that the lack of a discernible phenotype in Tmprss2 −/− mice suggests functional redundancy involving one or more of the type II transmembrane serine protease family members or other serine proteases. Alternatively, TMPRSS2 may contribute a specialized but nonvital function that is apparent only in the context of stress, disease, or other systemic perturbation.

TMPRSS4 is a type II transmembrane serine protease involved in cancer and viral infections

2012 ◽  
Vol 393 (9) ◽  
pp. 907-914 ◽  
Author(s):  
Anke Ohler ◽  
Christoph Becker-Pauly
Keyword(s):  
Serine Protease ◽  
Serine Proteases ◽  
Viral Infections ◽  
Proteolytic Enzymes ◽  
Metastatic Potential ◽  
Influenza Viruses ◽  
Type Ii ◽  
Serine Protease Family ◽  
Transmembrane Serine Protease ◽  
Almost All

Abstract Proteolytic enzymes are involved in almost all biological processes reflecting their importance in health and disease. The human genome contains nearly 600 protease-encoding genes forming more than 2% of the total human proteome. The serine proteases, with about 180 members, built the oldest and second largest family of human proteases. Ten years ago, a novel serine protease family named the type II transmembrane family (TTSP) was identified. This minireview summarizes the up-to-date knowledge about the still growing TTSPs, particularly focusing on the pathophysiological functions of the family member type II transmembrane serine protease (TMPRSS) 4. Recent studies provided important data on TMPRSS4 activity associated with the spreading of influenza viruses, mediated by the cleavage of hemagglutinin. Progression and metastatic potential of several cancers is concordant with an increased expression of TMPRSS4, though being a possible diagnostic marker. However, to benefit from TMPRSS4 as a therapeutic target, more data concerning its physiological relevance are needed, as done by a specific morpholino knockdown in zebrafish embryos.

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.

Discovery and Characterization Of a Highly Specific Antibody Inhibitor Of Plasma Kallikrein

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 1067-1067
Author(s):  
Jon A Kenniston ◽  
Daniel J Sexton ◽  
Diana Martik ◽  
Ryan R Faucette ◽  
Malini Viswanathan ◽  
...  
Keyword(s):  
Serine Protease ◽  
Serine Proteases ◽  
Therapeutic Option ◽  
Active Form ◽  
Crystallographic Structure ◽  
Human Antibody ◽  
Plasma Kallikrein ◽  
Related Proteins

Abstract The plasma-kallikrein kinin (contact) system contributes to the physiological and pathophysiological reactions of vascular biology. Activation of this pathway causes the release of the potent nonapeptide vasodilator bradykinin following proteolytic cleavage of high-molecular weight kininogen (HMWK) by the serine protease plasma kallikrein (pKal). Normal vascular homeostasis requires regulation of pKal activity by interactions with the C1-inhibitor (C1-INH). This is most apparent in individuals with hereditary angioedema (HAE), a disease characterized by a genetic deficiency in C1-INH that results in persistent pKal activity and consequent bradykinin release. These events can ultimately manifest as unpredictable and potentially fatal attacks of subcutaneous and mucosal edema. Inhibition of pKal proteolytic activity has proven to be a viable therapeutic option for HAE, however there remains an unmet medical need for a long-lasting prophylactic treatment for this disease. Given the potential for target specificity and long serum half-life with antibody therapeutics, we used phage display to select a fully human antibody inhibitor (DX-2930) specific for pKal. In vitro enzyme inhibition and affinity assays demonstrate that DX-2930 is a potent antibody inhibitor of pKal (Ki = 125 pM) that binds the active form of pKal, but not the proenzyme form (prekallikrein) or any other serine protease tested. DX-2930 binding consequently prohibits pKal from cleaving bradykinin out of HMWK and thereby prevents the activation of the bradykinin receptor B2. A 2.1Å resolution X-ray crystallographic structure of pKal complexed to a DX-2930 Fab construct supports these findings, demonstrating that the pKal proteolytic active site is intimately bound - and thereby occluded - by the Fab. This structural analysis provides both a rationale for the potency and specificity of DX-2930, and demonstrates the utility of using antibodies to specifically target an antigen among a family of related proteins (e.g. serine proteases). To further address the functional activity of DX-2930, we demonstrate that subcutaneous dosing of DX-2930 effectively reduces carrageenan-induced paw edema in vivo in rats when injected 24 hours prior to challenge. Combined with our finding that DX-2930 has a prolonged serum residence time in cynomolgus monkeys (t1/2 = 301 hours, SC), the data presented here demonstrates the potential of DX-2930 for the prophylactic inhibition of pKal-mediated diseases, such as HAE. Disclosures: Kenniston: Dyax Corp: Employment. Sexton:Dyax Corp: Employment. Martik:Dyax Corp: Employment, former employee of Dyax Corp Other. Faucette:Dyax Corp: Employment. Viswanathan:Dyax Corp: Employment. Kastrapeli:Dyax: Employment. Kopacz:Dyax Corp: Employment. Conley:Dyax Corp: Employment. Lindberg:Dyax Corp: Employment. Cosic:Dyax Corp: Employment. Comeau:Dyax Corp: Employment. Mason:Dyax Corp: Employment. DiLeo:Dyax Corp: Employment. Chen:Dyax Corp: Employment. Ladner:Dyax Corp: Employment. Edwards:Emerald Biostructures: Employment. TenHoor:Dyax Corp: Employment. Nixon:Dyax Corp: Employment. Adelman:Dyax Corp: Employment.

In vivo contribution of serine proteases to the proteolytic activation of γENaC in aldosterone-infused rats

2012 ◽  
Vol 303 (7) ◽  
pp. F939-F943 ◽  
Author(s):  
Kohei Uchimura ◽  
Yutaka Kakizoe ◽  
Tomoaki Onoue ◽  
Manabu Hayata ◽  
Jun Morinaga ◽  
...  
Keyword(s):  
Protease Inhibitor ◽  
Serine Protease ◽  
Serine Proteases ◽  
Serine Protease Inhibitor ◽  
Proteolytic Activation ◽  
New Strategy ◽  
Salt Sensitive Hypertension ◽  
Primary Cleavage

Aldosterone plays an important role in the regulation of blood pressure by modulating the activity of the epithelial sodium channel (ENaC) that consists of α-, β-, and γ-subunits. Aldosterone induces a molecular weight shift of γENaC from 85 to 70 kDa that is necessary for the channel activation. In vitro experiments demonstrated that a dual cleavage mechanism is responsible for this shift. It has been postulated that furin executes the primary cleavage in the Golgi and that the second cleavage is provided by other serine proteases such as prostasin or plasmin at the plasma membrane. However, the in vivo contribution of serine proteases to this cleavage remains unclear. To address this issue, we administered the synthetic serine protease inhibitor camostat mesilate (CM) to aldosterone-infused rats. CM decreased the abundance of the 70-kDa form of ENaC and led to a new 75-kDa form with a concomitant increase in the urinary Na-to-K ratio. Because CM inhibits the protease activity of serine proteases such as prostasin and plasmin, but not furin, our findings strongly indicate that CM inhibited the second cleavage of γENaC and subsequently suppressed ENaC activity. The results of our current studies also suggest the possibility that the synthetic serine protease inhibitor CM might represent a new strategy for the treatment of salt-sensitive hypertension in humans.

scholarly journals TMPRSS2 structure-phylogeny repositions Avoralstat for SARS-CoV-2 prophylaxis in mice

2021 ◽  
Author(s):  
Young Joo Sun ◽  
Gabriel Velez ◽  
Dylan Parsons ◽  
Kun Li ◽  
Miguel Ortiz ◽  
...  
Keyword(s):  
Viral Entry ◽  
Serine Proteases ◽  
Tertiary Structure ◽  
Airway Epithelial Cells ◽  
Airway Epithelial ◽  
Cell Infection ◽  
Effective Inhibition ◽  
Transmembrane Serine Protease

Drugs targeting host proteins can act prophylactically to reduce viral burden early in disease and limit morbidity, even with antivirals and vaccination. Transmembrane serine protease 2 (TMPRSS2) is a human protease required for SARS-CoV-2 viral entry and may represent such a target. We hypothesized drugs selected from proteins related by their tertiary structure, rather than their primary structure, were likely to interact with TMPRSS2. We created a structure-based phylogenetic computational tool 3DPhyloFold to systematically identify structurally similar serine proteases with known therapeutic inhibitors and demonstrated effective inhibition of SARS-CoV-2 infection in vitro and in vivo. Several candidate compounds, Avoralstat, PCI-27483, Antipain, and Soybean-Trypsin-Inhibitor, inhibited TMPRSS2 in biochemical and cell infection assays. Avoralstat, a clinically tested Kallikrein-related B1 inhibitor, inhibited SARS-CoV-2 entry and replication in human airway epithelial cells. In an in vivo proof of principle, Avoralstat significantly reduced lung tissue titers and mitigated weight-loss when administered prophylactically to SARS-CoV-2 susceptible mice indicating its potential to be repositioned for COVID-19 prophylaxis in humans.

Genetic Interactions Between the RhoA and Stubble-stubbloid Loci Suggest a Role for a Type II Transmembrane Serine Protease in Intracellular Signaling During Drosophila Imaginal Disc Morphogenesis

Genetics ◽  
2003 ◽  
Vol 165 (3) ◽  
pp. 1417-1432 ◽  
Author(s):  
Cynthia A Bayer ◽  
Susan R Halsell ◽  
James W Fristrom ◽  
Daniel P Kiehart ◽  
Laurence von Kalm
Keyword(s):  
Serine Protease ◽  
Signaling Pathway ◽  
Intracellular Signaling ◽  
Serine Proteases ◽  
Imaginal Discs ◽  
Type Ii ◽  
Gene Encoding ◽  
Developmental Abnormalities ◽  
Transmembrane Serine Protease ◽  
Rhoa Signaling

Abstract The Drosophila RhoA (Rho1) GTPase is essential for postembryonic morphogenesis of leg and wing imaginal discs. Mutations in RhoA enhance leg and wing defects associated with mutations in zipper, the gene encoding the heavy chain of nonmuscle myosin II. We demonstrate here that mutations affecting the RhoA signaling pathway also interact genetically with mutations in the Stubble-stubbloid (Sb-sbd) locus that encodes an unusual type II transmembrane serine protease required for normal leg and wing morphogenesis. In addition, a leg malformation phenotype associated with overexpression of Sb-sbd in prepupal leg discs is suppressed when RhoA gene dose is reduced, suggesting that RhoA and Sb-sbd act in a common pathway during leg morphogenesis. We also characterized six mutations identified as enhancers of zipper mutant leg defects. Three of these genes encode known members of the RhoA signaling pathway (RhoA, DRhoGEF2, and zipper). The remaining three enhancer of zipper mutations interact genetically with both RhoA and Sb-sbd mutations, suggesting that they encode additional components of the RhoA signaling pathway in imaginal discs. Our results provide evidence that the type II transmembrane serine proteases, a class of proteins linked to human developmental abnormalities and pathology, may be associated with intracellular signaling required for normal development.

BETULINIC ACID, A SUBSTANCE MUCH PROSPECTS IN MEDICINE

2011 ◽  
pp. 64-69
Author(s):  
Thi Hoai Nguyen ◽  
Thi Ni Na Le
Keyword(s):  
Medicinal Plants ◽  
Betulinic Acid ◽  
Structural Changes ◽  
Biological Activities ◽  
Diabetes Type ◽  
Type Ii ◽  
Diabetes Type Ii

Betulinic acid compounds derived from medicinal plants are attracting the attention of scientists with many important biological activities. Summary of the study showed that this compound and some of its derivatives work well in various in vitro and in vivo in the treatment of cancer, HIV, malaria and diabetes type II. The structural changes based on natural triterpen also noted the desire to find compounds that have powerful effects and safety.

Evidence that cell surface localization of serine protease activity facilitates cleavage of the protease activated receptor CDCP1

2018 ◽  
Vol 399 (9) ◽  
pp. 1091-1097
Author(s):  
Yaowu He ◽  
Janet C. Reid ◽  
Hui He ◽  
Brittney S. Harrington ◽  
Brittney Finlayson ◽  
...  
Keyword(s):  
Serine Protease ◽  
Protease Activity ◽  
Serine Proteases ◽  
Membrane Localization ◽  
Cub Domain ◽  
Serine Protease Activity ◽  
Surface Localization ◽  
Cell Surface Localization

Abstract The cellular receptor CUB domain containing protein 1 (CDCP1) is commonly elevated and functionally important in a range of cancers. CDCP1 is cleaved by serine proteases at adjacent sites, arginine 368 (R368) and lysine 369 (K369), which induces cell migration in vitro and metastasis in vivo. We demonstrate that membrane localization of serine protease activity increases efficacy of cleavage of CDCP1, and that both secreted and membrane anchored serine proteases can have distinct preferences for cleaving at CDCP1-R368 and CDCP1-K369. Approaches that disrupt membrane localization of CDCP1 cleaving serine proteases may interfere with the cancer promoting effects of CDCP1 proteolysis.

Evaluation of the In Vivo Acute Toxicity and In Vitro Hemolytic and Immunomodulatory Activities of the Moringa oleifera Flower Trypsin Inhibitor (MoFTI)

2020 ◽  
Vol 27 ◽  
Author(s):  
Leydianne Leite de Siqueira Patriota ◽  
Dayane Kelly Dias do Nascimento Santos ◽  
Bárbara Rafaela da Silva Barros ◽  
Lethícia Maria de Souza Aguiar ◽  
Yasmym Araújo Silva ◽  
...  
Keyword(s):  
Acute Toxicity ◽  
Trypsin Inhibitor ◽  
Hemolytic Activity ◽  
Moringa Oleifera ◽  
Biological Activities ◽  
Hematological Parameters ◽  
Behavioral Alterations ◽  
Female Mice

Background: Protease inhibitors have been isolated from plants and present several biological activities, including immunomod-ulatory action. Objective: This work aimed to evaluate a Moringa oleifera flower trypsin inhibitor (MoFTI) for acute toxicity in mice, hemolytic activity on mice erythrocytes and immunomodulatory effects on mice splenocytes. Methods: The acute toxicity was evaluated using Swiss female mice that received a single dose of the vehicle control or MoFTI (300 mg/kg, i.p.). Behavioral alterations were observed 15–240 min after administration, and survival, weight gain, and water and food consumption were analyzed daily. Organ weights and hematological parameters were analyzed after 14 days. Hemolytic activity of MoFTI was tested using Swiss female mice erythrocytes. Splenocytes obtained from BALB/c mice were cultured in the absence or presence of MoFTI for the evaluation of cell viability and proliferation. Mitochondrial membrane potential (ΔΨm) and reactive oxygen species (ROS) levels were also determined. Furthermore, the culture supernatants were analyzed for the presence of cytokines and nitric oxide (NO). Results: MoFTI did not cause death or any adverse effects on the mice except for abdominal contortions at 15–30 min after administration. MoFTI did not exhibit a significant hemolytic effect. In addition, MoFTI did not induce apoptosis or necrosis in splenocytes and had no effect on cell proliferation. Increases in cytosolic and mitochondrial ROS release, as well as ΔΨm reduction, were observed in MoFTI-treated cells. MoFTI was observed to induce TNF-α, IFN-γ, IL-6, IL-10, and NO release. Conclusion: These results contribute to the ongoing evaluation of the antitumor potential of MoFTI and its effects on other immunological targets.

Metabolomics of Healthy Berry Fruits

2019 ◽  
Vol 25 (37) ◽  
pp. 4888-4902 ◽  
Author(s):  
Gilda D'Urso ◽  
Sonia Piacente ◽  
Cosimo Pizza ◽  
Paola Montoro
Keyword(s):  
Biological Activities ◽  
Biologically Active ◽  
In Vivo Studies ◽  
Bioactive Metabolites ◽  
Active Metabolites ◽  
Positive Effects ◽  
Cell Functions ◽  
Berry Fruits

The consumption of berry-type fruits has become very popular in recent years because of their positive effects on human health. Berries are in fact widely known for their health-promoting benefits, including prevention of chronic disease, cardiovascular disease and cancer. Berries are a rich source of bioactive metabolites, such as vitamins, minerals, and phenolic compounds, mainly anthocyanins. Numerous in vitro and in vivo studies recognized the health effects of berries and their function as bioactive modulators of various cell functions associated with oxidative stress. Plants have one of the largest metabolome databases, with over 1200 papers on plant metabolomics published only in the last decade. Mass spectrometry (MS) and NMR (Nuclear Magnetic Resonance) are the most important analytical technologies on which the emerging ''omics'' approaches are based. They may provide detection and quantization of thousands of biologically active metabolites from a tissue, working in a ''global'' or ''targeted'' manner, down to ultra-trace levels. In the present review, we highlighted the use of MS and NMR-based strategies and Multivariate Data Analysis for the valorization of berries known for their biological activities, important as food and often used in the preparation of nutraceutical formulations.

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