scholarly journals In vivo and in vitro Activity of 1-aminocyclopropane-1-carboxylic Acid Oxidase in Germinating Seeds of China Aster (Callistephus chinensis Nees)

2020 ◽  
Vol 28 (2) ◽  
pp. 11-20
Author(s):  
Mariusz Chojnowski ◽  
Anna Skorupińska
Keyword(s):  
Carboxylic Acid ◽  
Active Sites ◽  
Sensu Stricto ◽  
Saturation Level ◽  
Acid Oxidase ◽  
Enzymatic Extract ◽  
Acc Conversion ◽  
Germinating Seeds

AbstractThe activity of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO; EC 1.4.3.3) in germinating seeds of Callistephus chinensis was studied. For maximum recovery of ACO activity in vitro, the presence of 10% (w/v) insoluble polyvinylpolypyrrolidone (PVPP) and 30% of glycerol in the extraction medium was necessary. The optimum pH for this activity was 7.0. Ethylene production by whole achenes or enzymatic extract increased due to increasing 1-aminocyclopropane-1-carboxylic acid (ACC) concentrations. Saturation level of ACC for in vivo ACO activity was 10−1 M and Vmax was 10.89 nL C2H4·mg protein−1·h−1. For in vitro ACO activity, the saturation level of ACC was 10−3 M and Vmax was 2.299 nL C2H4·mg protein−1·h−1. Both, in vivo and in vitro ACO activities did not follow Michaelis-Menten kinetics. The Hill coefficients (h) were estimated on the basis of non-linear estimation. Their values were 0.63 for in vivo ACO activity and 1.73 for in vitro ACO activity. The experimental data show that ACO from C. chinensis seeds is an oligomeric enzyme with at least two active sites. During seed germination, in vitro ACO activity was detectable after 12 hours of imbibition, while in vivo ACC conversion to ethylene was observed after 24 h, i.e. – after radicle protrusion. The activity of ACO in C. chinensis seeds is associated with germination sensu stricto, and might be a good marker of this process.

Action of Thioglycosides of 1,2,4-Triazoles and Imidazoles on the Oxidative Stress and Glycosidases in Mice with Molecular Docking

2019 ◽  
Vol 16 (6) ◽  
pp. 696-710
Author(s):  
Mahmoud Balbaa ◽  
Doaa Awad ◽  
Ahmad Abd Elaal ◽  
Shimaa Mahsoub ◽  
Mayssaa Moharram ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Molecular Docking ◽  
Active Sites ◽  
Biological Activities ◽  
Imidazole Ring ◽  
Quantitative Structure Activity Relationship ◽  
Binding Interaction ◽  
Qsar Study

Background: ,2,3-Triazoles and imidazoles are important five-membered heterocyclic scaffolds due to their extensive biological activities. These products have been an area of growing interest to many researchers around the world because of their enormous pharmaceutical scope. Methods: The in vivo and in vitro enzyme inhibition of some thioglycosides encompassing 1,2,4- triazole N1, N2, and N3 and/or imidazole moieties N4, N5, and N6. The effect on the antioxidant enzymes (superoxide dismutase, glutathione S-transferase, glutathione peroxidase and catalase) was investigated as well as their effect on α-glucosidase and β-glucuronidase. Molecular docking studies were carried out to investigate the mode of the binding interaction of the compounds with α- glucosidase and β -glucuronidase. In addition, quantitative structure-activity relationship (QSAR) investigation was applied to find out the correlation between toxicity and physicochemical properties. Results: The decrease of the antioxidant status was revealed by the in vivo effect of the tested compounds. Furthermore, the in vivo and in vitro inhibitory effects of the tested compounds were clearly pronounced on α-glucosidase, but not β-glucuronidase. The IC50 and Ki values revealed that the thioglycoside - based 1,2,4-triazole N3 possesses a high inhibitory action. In addition, the in vitro studies demonstrated that the whole tested 1,2,4-triazole are potent inhibitors with a Ki magnitude of 10-6 and exhibited a competitive type inhibition. On the other hand, the thioglycosides - based imidazole ring showed an antioxidant activity and exerted a slight in vivo stimulation of α-glucosidase and β- glucuronidase. Molecular docking proved that the compounds exhibited binding affinity with the active sites of α -glucosidase and β-glucuronidase (docking score ranged from -2.320 to -4.370 kcal/mol). Furthermore, QSAR study revealed that the HBD and RB were found to have an overall significant correlation with the toxicity. Conclusion: These data suggest that the inhibition of α-glucosidase is accompanied by an oxidative stress action.

scholarly journals Echinococcus granulosus sensu stricto: silencing of thioredoxin peroxidase impairs the differentiation of protoscoleces into metacestodes

Parasite ◽  
2018 ◽  
Vol 25 ◽  
pp. 57 ◽  
Author(s):  
Hui Wang ◽  
Jun Li ◽  
Chuanshan Zhang ◽  
Baoping Guo ◽  
Qin Wei ◽  
...  
Keyword(s):  
Echinococcus Granulosus ◽  
Sensu Stricto ◽  
Antioxidant Defence ◽  
Culture Studies ◽  
Antioxidant Defence System ◽  
Thioredoxin Peroxidase ◽  
Survival And Growth ◽  
Echinococcus Granulosus Sensu Lato

Cystic echinococcosis (CE) is a cosmopolitan parasitic disease caused by infection with the larval stage of Echinococcus granulosus sensu lato. Thioredoxin peroxidase (TPx) may play an essential role in the antioxidant defence system of E. granulosus s.l. as neither catalase nor glutathione peroxidase activities have been detected in the parasite. However, it is not known whether TPx affects the survival and growth of E. granulosus s.l. during development. In this study, three fragments of siRNA specific for EgTPx (siRNA-1/2/3) were designed and transfected into protoscoleces of E. granulosus sensu stricto by electroporation. Quantitative real-time PCR and Western blotting analysis showed that siRNA-3 significantly reduced the expression of EgTPx. Coincidentally, knockdown of EgTPx expression in protoscoleces with siRNA-3 significantly reduced the viability of the parasite under oxidative stress induced by 0.6 mM H2O2. In vitro culture studies showed that protoscoleces treated with siRNA-3 reduced pre-microcyst formation. In vivo experiments showed that injecting mice intraperitoneally with protoscoleces treated with siRNA-3 resulted in a significant reduction in the number, size and weight of CE cysts compared with those of control animals. Silencing of EgTPx led to the impairment of growth of E. granulosus s.s. both in vitro and in vivo, indicating that EgTPx is an important factor for protoscoleces survival and plays an important role in the antioxidant defence against the host during development.

scholarly journals In Vitro and In Vivo Inhibition of the 2 Active Sites of ACE by Omapatrilat, a Vasopeptidase Inhibitor

Hypertension ◽  
2000 ◽  
Vol 35 (6) ◽  
pp. 1226-1231 ◽  
Author(s):  
Michel Azizi ◽  
Christine Massien ◽  
Annie Michaud ◽  
Pierre Corvol
Keyword(s):  
Active Sites ◽  
Vasopeptidase Inhibitor

TrioGEF1 controls Rac- and Cdc42-dependent cell structures through the direct activation of rhoG

2000 ◽  
Vol 113 (4) ◽  
pp. 729-739 ◽  
Author(s):  
A. Blangy ◽  
E. Vignal ◽  
S. Schmidt ◽  
A. Debant ◽  
C. Gauthier-Rouviere ◽  
...  
Keyword(s):  
Rho Gtpases ◽  
Active Sites ◽  
Dominant Negative ◽  
Signaling Cascade ◽  
Microtubule Network ◽  
Cell Structures ◽  
Dependent Cell

Rho GTPases regulate the morphology of cells stimulated by extracellular ligands. Their activation is controlled by guanine exchange factors (GEF) that catalyze their binding to GTP. The multidomain Trio protein represents an emerging class of Ρ regulators that contain two GEF domains of distinct specificities. We report here the characterization of Rho signaling pathways activated by the N-terminal GEF domain of Trio (TrioD1). In fibroblasts, TrioD1 triggers the formation of particular cell structures, similar to those elicited by RhoG, a GTPase known to activate both Rac1 and Cdc42Hs. In addition, the activity of TrioD1 requires the microtubule network and relocalizes RhoG at the active sites of the plasma membrane. Using a classical in vitro exchange assay, TrioD1 displays a higher GEF activity on RhoG than on Rac1. In fibroblasts, expression of dominant negative RhoG mutants totally abolished TrioD1 signaling, whereas dominant negative Rac1 and Cdc42Hs only led to partial and complementary inhibitions. Finally, expression of a Rho Binding Domain that specifically binds RhoG(GTP) led to the complete abolition of TrioD1 signaling, which strongly supports Rac1 not being activated by TrioD1 in vivo. These data demonstrate that Trio controls a signaling cascade that activates RhoG, which in turn activates Rac1 and Cdc42Hs.

scholarly journals Discovery of a dual inhibitor of NQO1 and GSTP1 for treating glioblastoma

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Kecheng Lei ◽  
Xiaoxia Gu ◽  
Alvaro G. Alvarado ◽  
Yuhong Du ◽  
Shilin Luo ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Crystal Structures ◽  
Active Sites ◽  
Redox Homeostasis ◽  
Growth Factor Receptor ◽  
Quinone Oxidoreductase ◽  
Cancer Proliferation ◽  
Dual Inhibitor

Abstract Background Glioblastoma (GBM) is a universally lethal tumor with frequently overexpressed or mutated epidermal growth factor receptor (EGFR). NADPH quinone oxidoreductase 1 (NQO1) and glutathione-S-transferase Pi 1 (GSTP1) are commonly upregulated in GBM. NQO1 and GSTP1 decrease the formation of reactive oxygen species (ROS), which mediates the oxidative stress and promotes GBM cell proliferation. Methods High-throughput screen was used for agents selectively active against GBM cells with EGFRvIII mutations. Co-crystal structures were revealed molecular details of target recognition. Pharmacological and gene knockdown/overexpression approaches were used to investigate the oxidative stress in vitro and in vivo. Results We identified a small molecular inhibitor, “MNPC,” that binds to both NQO1 and GSTP1 with high affinity and selectivity. MNPC inhibits NQO1 and GSTP1 enzymes and induces apoptosis in GBM, specifically inhibiting the growth of cell lines and primary GBM bearing the EGFRvIII mutation. Co-crystal structures between MNPC and NQO1, and molecular docking of MNPC with GSTP1 reveal that it binds the active sites and acts as a potent dual inhibitor. Inactivation of both NQO1 and GSTP1 with siRNA or MNPC results in imbalanced redox homeostasis, leading to apoptosis and mitigated cancer proliferation in vitro and in vivo. Conclusions Thus, MNPC, a dual inhibitor for both NQO1 and GSTP1, provides a novel lead compound for treating GBM via the exploitation of specific vulnerabilities created by mutant EGFR.

Fate of Safening Agent L-2-Oxothiazolidine-4-Carboxylic Acid in Sorghum (Sorghum bicolor)

Weed Science ◽  
1990 ◽  
Vol 38 (3) ◽  
pp. 201-205 ◽  
Author(s):  
James L. Hilton ◽  
Parthasarathy Pillai ◽  
Helen A. Norman
Keyword(s):  
Carboxylic Acid ◽  
Sorghum Bicolor ◽  
Culture Medium ◽  
In Vitro Assay ◽  
Thiol Compounds ◽  
Vitro Assay ◽  
Herbicide Safener ◽  
Liquid Culture Medium ◽  
Germinating Seeds

The herbicide safener OTC (L-2-oxothiazolidine-4-carboxylic acid) increased the amount of reduced thiol compounds in sorghum [Sorghum bicolor(L.) Moench. ‘DK 427′] seedlings. When seedlings were grown in liquid culture medium containing35S-OTC, the compound was metabolized to radiolabeled cysteine and glutathione. The addition of tridiphane [2-(3,5-dichlorophenyl)-2-(2,2,2-trichloroethyl)oxirane] increased conversion of35S-OTC to cysteine and resulted in the formation of one additional35S-labeled compound. When35S-glutathione was injected into germinating seeds it was converted to35S-cysteine and both thiols were subsequently found in roots and shoots. Seeds injected with35S-OTC both translocated the compound to developing roots and shoots and metabolized35S-OTC to cysteine and glutathione. Excised roots and shoots also metabolized35S-OTC to the thiols. In an in vitro assay the enzyme 5-oxoprolinase converted OTC to cysteine.

scholarly journals Vipera berus berus Venom from Russia: Venomics, Bioactivities and Preclinical Assessment of Microgen Antivenom

Toxins ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 90 ◽  
Author(s):  
Ruslan I. Al-Shekhadat ◽  
Ksenia S. Lopushanskaya ◽  
Álvaro Segura ◽  
José María Gutiérrez ◽  
Juan J. Calvete ◽  
...  
Keyword(s):  
Binding Capacity ◽  
Secretory Protein ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Vipera Berus ◽  
Phospholipases A2 ◽  
Toxin Binding ◽  
Main Effects

The common European adder, Vipera berus berus, is a medically relevant species, which is widely distributed in Russia and thus, is responsible for most snakebite accidents in Russia. We have investigated the toxic and enzymatic activities and have determined the proteomic composition of its venom. Phospholipases A2 (PLA2, 25.3% of the venom proteome), serine proteinases (SVSP, 16.2%), metalloproteinases (SVMP, 17.2%), vasoactive peptides (bradykinin-potentiating peptides (BPPs), 9.5% and C-type natriuretic peptides (C-NAP, 7.8%), cysteine-rich secretory protein (CRISP, 8%) and L-amino acid oxidase (LAO, 7.3%) represent the major toxin classes found in V. b. berus (Russia) venom. This study was also designed to assess the in vivo and in vitro preclinical efficacy of the Russian Microgen antivenom in neutralizing the main effects of V. b. berus venom. The results show that this antivenom is capable of neutralizing the lethal, hemorrhagic and PLA2 activities. Third-generation antivenomics was applied to quantify the toxin-recognition landscape and the maximal binding capacity of the antivenom for each component of the venom. The antivenomics analysis revealed that 6.24% of the anti-V. b. berus F(ab’)2 molecules fraction are toxin-binding antibodies, 60% of which represent clinically relevant antivenom molecules.

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