scholarly journals Identification of Potential Inhibitors of Calcium/Calmodulin-Dependent Protein Kinase IV from Bioactive Phytoconstituents

2020 ◽  
Vol 2020 ◽  
pp. 1-14 ◽  
Author(s):  
Preeti Gupta ◽  
Shama Khan ◽  
Zeynab Fakhar ◽  
Afzal Hussain ◽  
Md. Tabish Rehman ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Neurodegenerative Diseases ◽  
Simulation Analysis ◽  
Inhibition Mechanism ◽  
Binding Studies ◽  
Ligand Complex ◽  
Dependent Protein Kinase ◽  
Calcium Calmodulin Dependent ◽  
Dependent Protein

Calcium/calmodulin-dependent protein kinase IV (CaMKIV) is an upstream regulator of CaMKK-CaMKIV signaling cascade that activates various transcription factors, thereby regulating several cellular activities including, neuronal communication and immune response. Owing to the abnormal expression in cancer and neurodegenerative diseases, the CaMKIV has been considered a potential drug target. In the present study, we checked the binding affinity of plant-derived natural compounds viz., quercetin, ellagic acid (EA), simvastatin, capsaicin, ursolic acid, DL-α-tocopherol acetate, and limonin towards CaMKIV. Molecular docking and fluorescence binding studies showed that EA and quercetin bind to the CaMKIV with a considerable affinity in comparison to other compounds. Enzyme inhibition assay revealed that both EA and quercetin inhibit CaMKIV activity with their IC50 values in the micromolar range. To get atomistic insights into the mode of interactions, inhibition mechanism, and the stability of the CaMKIV-ligand complex, a 100 ns MD simulation analysis was performed. Both EA and quercetin bind to the catalytically important residues of active site pocket of CaMKIV forming enough stabilizing interactions presumably inhibiting enzyme activity. Moreover, no significant structural change in the CaMKIV was observed upon binding of EA and quercetin. In conclusion, this study illustrates the application of phytoconstituents in the development of therapeutic molecules targeting CaMKIV having implications in cancer and neurodegenerative diseases after in vivo validation.

scholarly journals Anti-echinococcal effect of verapamil involving the regulation of the calcium/calmodulin-dependent protein kinase II response in vitro and in a murine infection model

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Hai-Jun Gao ◽  
Xu-Dong Sun ◽  
Yan-Ping Luo ◽  
Hua-Sheng Pang ◽  
Xing-Ming Ma ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Mrna Expression ◽  
Echinococcus Granulosus ◽  
Calcium Content ◽  
Mrna Levels ◽  
Dependent Protein Kinase ◽  
Calcium Calmodulin Dependent ◽  
Dependent Protein

Abstract Background Echinococcosis, which is caused by the larvae of cestodes of the genus Echinococcus, is a parasitic zoonosis that poses a serious threat to the health of humans and animals globally. Albendazole is the drug of choice for the treatment of echinococcosis, but it is difficult to meet clinical goals with this chemotherapy due to its low cure rate and associated side effects after its long-term use. Hence, novel anti-parasitic targets and effective treatment alternatives are urgently needed. A previous study showed that verapamil (Vepm) can suppress the growth of Echinococcus granulosus larvae; however, the mechanism of this effect remains unclear. The aim of the present study was to gain insight into the anti-echinococcal effect of Vepm on Echinococcus with a particular focus on the regulatory effect of Vepm on calcium/calmodulin-dependent protein kinase II (Ca2+/CaM-CaMKII) in infected mice. Methods The anti-echinococcal effects of Vepm on Echinococcus granulosus protoscoleces (PSC) in vitro and Echinococcus multilocularis metacestodes in infected mice were assessed. The morphological alterations in Echinococcus spp. induced by Vepm were observed by scanning electron microscopy (SEM), and the changes in calcium content in both the parasite and mouse serum and liver were measured by SEM-energy dispersive spectrometry, inductively coupled plasma mass spectrometry and alizarin red staining. Additionally, the changes in the protein and mRNA levels of CaM and CaMKII in infected mice, and in the mRNA levels of CaMKII in E. granulosus PSC, were evaluated after treatment with Vepm by immunohistochemistry and/or real-time quantitative polymerase chain reaction. Results In vitro, E. granulosus PSC could be killed by Vepm at a concentration of 0.5 μg/ml or higher within 8 days. Under these conditions, the ultrastructure of PSC was damaged, and this damage was accompanied by obvious calcium loss and downregulation of CaMKII mRNA expression. In vivo, the weight and the calcium content of E. multilocularis metacestodes from mice were reduced after treatment with 40 mg/kg Vepm, and an elevation of the calcium content in the sera and livers of infected mice was observed. In addition, downregulation of CaM and CaMKII protein and mRNA expression in the livers of mice infected with E. multilocularis metacestodes was found after treatment with Vepm. Conclusions Vepm exerted a parasiticidal effect against Echinococcus both in vitro and in vivo through downregulating the expression of Ca2+/CaM-CaMKII, which was over-activated by parasitic infection. The results suggest that Ca2+/CaM-CaMKII may be a novel drug target, and that Vepm is a potential anti-echinococcal drug for the future control of echinococcosis.

scholarly journals A Molecular Mechanism of Integrin Crosstalk: αvβ3 Suppression of Calcium/Calmodulin-dependent Protein Kinase II Regulates α5β1 Function

1999 ◽  
Vol 145 (4) ◽  
pp. 889-897 ◽  
Author(s):  
Scott D. Blystone ◽  
Suzanne E. Slater ◽  
Matthew P. Williams ◽  
Michael T. Crow ◽  
Eric J. Brown
Keyword(s):  
Protein Kinase ◽  
Function Analysis ◽  
Cytoplasmic Tail ◽  
Integrin Αvβ3 ◽  
Dependent Protein Kinase ◽  
Calcium Calmodulin Dependent ◽  
Protein Kinase Ii ◽  
Dependent Protein ◽  
And Migration

Many cells express more than one integrin receptor for extracellular matrix, and in vivo these receptors may be simultaneously engaged. Ligation of one integrin may influence the behavior of others on the cell, a phenomenon we have called integrin crosstalk. Ligation of the integrin αvβ3 inhibits both phagocytosis and migration mediated by α5β1 on the same cell, and the β3 cytoplasmic tail is necessary and sufficient for this regulation of α5β1. Ligation of α5β1 activates the calcium- and calmodulin-dependent protein kinase II (CamKII). This activation is required for α5β1-mediated phagocytosis and migration. Simultaneous ligation of αvβ3 or expression of a chimeric molecule with a free β3 cytoplasmic tail prevents α5β1-mediated activation of CamKII. Expression of a constitutively active CamKII restores α5β1 functions blocked by αvβ3-initiated integrin crosstalk. Thus, αvβ3 inhibition of α5β1 activation of CamKII is required for its role in integrin crosstalk. Structure-function analysis of the β3 cytoplasmic tail demonstrates a requirement for Ser752 in β3-mediated suppression of CamKII activation, while crosstalk is independent of Tyr747 and Tyr759, implicating Ser752, but not β3 tyrosine phosphorylation in initiation of the αvβ3 signal for integrin crosstalk.

scholarly journals Anti-echinococcal effect of verapamil involving the regulation of the calcium/calmodulin-dependent protein kinase Ⅱ response in vitro and in a murine infection model

2021 ◽  
Author(s):  
Hai-Jun Gao ◽  
Xu-Dong Sun ◽  
Yan-Ping Luo ◽  
Hua-Sheng Pang ◽  
Xing-Ming Ma ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Mrna Expression ◽  
Calcium Content ◽  
Mouse Serum ◽  
Mrna Levels ◽  
Dependent Protein Kinase ◽  
Calcium Calmodulin Dependent ◽  
Dependent Protein

Abstract Background: Echinococcosis caused by the larvae of cestodes of the genus Echinococcus is a parasitic zoonosis that poses a serious threat to the health of humans and animals globally. Albendazole is the drug of choice for the treatment of echinococcosis, but it is difficult to meet clinical goals with this chemotherapy due to its low cure rate and the assocaiated side effects after long-time use. Hence, novel anti-parasitic targets and effective treatment alternatives are urgently needed. A previous study showed that verapamil can suppress the growth of Echinococcus granulosus larvae; however, the mechanism of this effect remains unclear. The aim of the present study was to gain insight into the anti-echinococcal effect of verapamil on Echinococcus with a particular focus on the regulatory effect of verapamil on calcium/calmodulin-dependent protein kinase Ⅱ (Ca2+/CaM-CamKⅡ) in infected mice.Methods: The anti-echinococcal effects of verapamil on E. granulosus protoscoleces (PSC) in vitro and E. multilocularis metacestodes in infected mice were assessed. The morphological alterations in Echinococcus spp. induced by verapamil were observed by scanning electron microscopy (SEM), and the changes in calcium content in both the parasite and mouse serum and liver were measured by SEM-energy dispersive spectrometry, inductively coupled plasma mass spectrometry and Alizarin red staining. Additionally, the changes in the protein and mRNA levels of CaM and CamKⅡ in infected mice, and in the mRNA levels of CamKⅡ in E. granulosus PSC were evaluated after treatment with verapamil by immunohistochemistry and/or real-time quantitative polymerase chain reaction.Results: In vitro, E. granulosus PSC could be killed by verapamil at a concentration of 0.5 μg/mL or higher within 8 days. Under these conditions, the ultrastructure of PSC was damaged, and this damage was accompanied obvious calcium loss and downregulation of CamKⅡ mRNA expression. In vivo, the weight and the calcium content of E. multilocularis metacestodes from mice were reduced after treatment with 40 mg/kg verapamil, and an elevation of the calcium content in the sera and livers of infected mice was observed. In addition, downregulation of CaM and CamKⅡ protein and mRNA expression in the livers of mice infected with E. multilocularis metacestodes was found after treatment with verapamil.Conclusions: Verapamil exerted a parasiticidal effect against Echinococcus both in vitro and in vivo through downregulating the expression of Ca2+/CaM-CamKⅡ, which were over-activated by parasitic infection. The results suggested that Ca2+/CaM-CamKⅡ may be a novel drug-target, and verapamil was shown to be a potential anti-echinococcal drug for controlling echinococcosis in the future.

scholarly journals Anti-echinococcal effect of verapamil involving regulating calcium/calmodulin-dependent protein kinase Ⅱ response in vitro and in a murine infection model

2020 ◽  
Author(s):  
Hai-Jun Gao ◽  
Xu-Dong Sun ◽  
Yan-Ping Luo ◽  
Hua-Sheng Pang ◽  
Xing-Ming Ma ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Calcium Content ◽  
Morphological Alteration ◽  
Mrna Levels ◽  
Down Regulation ◽  
Dependent Protein Kinase ◽  
Calcium Calmodulin Dependent ◽  
Dependent Protein

Abstract Background: Echinococcosis caused by the larval stage of cestode of the genus Echinococcus is a parasitic zoonosis, imposing serious threat on the health of humans and animals globally. Albendazole is the drug of choice for treatment of echinococcosis, but it is difficult to meet the clinical challenge in chemotherapy due to its low curative rate and severe side effects. Hence, novel anti-parasitic targets and effective treatment alternatives are urgently needed. Previous study has showed that verapamil can suppress the growth of Echinococcus granulosus (E. granulosus) larva, but the mechanism remains unclear. The aim of the present study is to gain insight into the anti-echinococcal effect of verapamil on Echinococcus with particular focus on the regulatory role of verapamil to calcium/calmodulin-dependent protein kinase Ⅱ (Ca2+/CaM-CamK Ⅱ) in infected mice.Methods: The anti-echinococcal effect of verapamil on E. granulosus protoscolex (PSCx) in vitro and Echinococcus multilocularis (E. multilocularis) metacestodes in infected mice were assessed. The morphological alteration of Echinococcus spp. induced by verapamil was observed by scanning electron microscope (SEM), and the changes of calcium content in both parasite and mice sera and livers were measured by SEM-energy dispersive spectrometer, inductively coupled plasma mass spectrometry and alizarin red staining accordingly. Additionally, the changes on the protein and mRNA levels of CaM and CamK Ⅱ in infected mice, and on the mRNA levels of CamK Ⅱ in E. granulosus PSCx after treatment of verapamil were evaluated by immunohistochemistry or/and real-time quantitative polymerase chain reaction (RT-qPCR).Results: In vitro, E. granulosus PSCx could be killed by verapamil at 0.5 μg/mL or more within 8 days. Under these conditions, the ultrastructure of PSCx was damaged and accompanied with obvious calcium loss and down-regulation of CamK Ⅱ mRNA. In vivo, the weight and the calcium content of E. multilocularis metacestodes from the mice were reduced after treatment with verapamil at 40 mg/kg, meanwhile, an elevation of calcium content in the serum and liver of infected mice was observed. In addition, down-regulation of both the over-expressed CaM and CamK Ⅱ proteins and mRNAs in the liver of mice infected with E. multilocularis metacestodes were found after treatment of verapamil.Conclusions: Verapamil exerted a parasiticidal activity on Echinococcus both in vitro and in vivo through down-regulation of Ca2+/CaM-CamK Ⅱ that was over-activated by parasitic infection. It has been speculated that the Ca2+/CaM-CamK Ⅱ can be a novel drug-target and verapamil is shown to be a potential anti-echinococcal drug for controlling echinococcosis in the future.

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