12,352 results on '"Kinase activity"'
Search Results
2. Single molecule array measures of LRRK2 kinase activity in serum link Parkinson’s disease severity to peripheral inflammation
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Yuan, Yuan, Li, Huizhong, Sreeram, Kashyap, Malankhanova, Tuyana, Boddu, Ravindra, Strader, Samuel, Chang, Allison, Bryant, Nicole, Yacoubian, Talene A., Standaert, David G., Erb, Madalynn, Moore, Darren J., Sanders, Laurie H., Lutz, Michael W., Velmeshev, Dmitry, and West, Andrew B.
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- 2024
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3. LRRK2 is involved in the chemotaxis of neutrophils and differentiated HL-60 cells, and the inhibition of LRRK2 kinase activity increases fMLP-induced chemotactic activity
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Mazaki, Yuichi, Handa, Haruka, Fumoto, Yoshizuki, Horinouchi, Takahiro, and Onodera, Yasuhito
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- 2023
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4. Potential selection for lipid kinase activity and spermatogenesis in Henan native pig breeds and growth shaping by introgression of European genes
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Qiao, Ruimin, Li, Xinjian, Madsen, Ole, Groenen, Martien A. M., Xu, Pan, Wang, Kejun, Han, Xuelei, Li, Gaiying, Li, Xiuling, and Li, Kui
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- 2023
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5. ASFV pD345L protein negatively regulates NF-κB signalling by inhibiting IKK kinase activity
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Chen, Huan, Wang, Zhenzhong, Gao, Xiaoyu, Lv, Jiaxuan, Hu, Yongxin, Jung, Yong-Sam, Zhu, Shanyuan, Wu, Xiaodong, Qian, Yingjuan, and Dai, Jianjun
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- 2022
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6. Impairments in SHMT2 expression or cellular folate availability reduce oxidative phosphorylation and pyruvate kinase activity
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Fiddler, Joanna L., Blum, Jamie E., Heyden, Katarina E., Castillo, Luisa F., Thalacker-Mercer, Anna E., and Field, Martha S.
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- 2023
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7. The ketogenic diet increases Neuregulin 1 expression via elevating histone acetylation and its anti-seizure effect requires ErbB4 kinase activity
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Wang, Jin, Huang, Jie, Yao, Shan, Wu, Jia-Hui, Li, Hui-Bin, Gao, Feng, Wang, Ying, Huang, Guo-Bin, You, Qiang-Long, Li, Jianhua, Chen, Xiaohui, and Sun, Xiang-Dong
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- 2021
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8. Inference of kinase-signaling networks in human myeloid cell line models by Phosphoproteomics using kinase activity enrichment analysis (KAEA)
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Hallal, Mahmoud, Braga-Lagache, Sophie, Jankovic, Jovana, Simillion, Cedric, Bruggmann, Rémy, Uldry, Anne-Christine, Allam, Ramanjaneyulu, Heller, Manfred, and Bonadies, Nicolas
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- 2021
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9. Both levoglucosan kinase activity and transport capacity limit the utilization of levoglucosan in Saccharomyces cerevisiae
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Yang, Mengdan, Wei, Tiandi, Wang, Kai, Jiang, Liqun, Zeng, Dihao, Sun, Xinhua, Liu, Weifeng, and Shen, Yu
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- 2022
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10. Time dependent effect of cold ischemia on the phosphoproteome and protein kinase activity in fresh-frozen colorectal cancer tissue obtained from patients
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Buffart, Tineke E., van den Oord, Rosanne A. H. M., van den Berg, Adriënne, Hilhorst, Riet, Bastiaensen, Niek, Pruijt, Hans F. M., van den Brule, Adriaan, Nooijen, Peet, Labots, Mariette, de Goeij-de Haas, Richard R., Dekker, Henk, Piersma, Sander R., Pham, Thang V., van der Leij, Theo, de Wijn, Rik, Ruijtenbeek, Rob, Jiménez, Connie R., and Verheul, Henk M. W.
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- 2021
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11. Inhibition of LRRK2 kinase activity promotes anterograde axonal transport and presynaptic targeting of α-synuclein
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Brzozowski, Charlotte F., Hijaz, Baraa A., Singh, Vijay, Gcwensa, Nolwazi Z., Kelly, Kaela, Boyden, Edward S., West, Andrew B., Sarkar, Deblina, and Volpicelli-Daley, Laura A.
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- 2021
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12. SPOP-mediated ubiquitination and degradation of PDK1 suppresses AKT kinase activity and oncogenic functions
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Jiang, Qiwei, Zheng, Nana, Bu, Lang, Zhang, Xiaomei, Zhang, Xiaoling, Wu, Yuanzhong, Su, Yaqing, Wang, Lei, Zhang, Xiaomin, Ren, Shancheng, Dai, Xiangpeng, Wu, Depei, Xie, Wei, Wei, Wenyi, Zhu, Yasheng, and Guo, Jianping
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- 2021
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13. Correction to: Inhaled gold nanoparticles cause cerebral edema and upregulate endothelial aquaporin 1 expression, involving caveolin 1 dependent repression of extracellular regulated protein kinase activity
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Chen, Ching-Yi, Liao, Po-Lin, Tsai, Chi-Hao, Chan, Yen-Ju, Cheng, Yu-Wen, Hwang, Ling-Ling, Lin, Kuan-Hung, Yen, Ting-Ling, and Li, Ching-Hao
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- 2019
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14. Inhaled gold nanoparticles cause cerebral edema and upregulate endothelial aquaporin 1 expression, involving caveolin 1 dependent repression of extracellular regulated protein kinase activity
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Chen, Ching-Yi, Liao, Po-Lin, Tsai, Chi-Hao, Chan, Yen-Ju, Cheng, Yu-Wen, Hwang, Ling-Ling, Lin, Kuan-Hung, Yen, Ting-Ling, and Li, Ching-Hao
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- 2019
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15. Neurodegeneration in SCA14 is associated with increased PKCγ kinase activity, mislocalization and aggregation
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Wong, Maggie M. K., Hoekstra, Stephanie D., Vowles, Jane, Watson, Lauren M., Fuller, Geraint, Németh, Andrea H., Cowley, Sally A., Ansorge, Olaf, Talbot, Kevin, and Becker, Esther B. E.
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- 2018
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16. Nefopam downregulates autophagy and c-Jun N-terminal kinase activity in the regulation of neuropathic pain development following spinal nerve ligation
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Oh, Seon-Hee, Yoon, Myung Ha, Lim, Kyung Joon, Yu, Byung Sik, Jee, In Gook, and Jung, Ki Tae
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- 2018
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17. AKT1 restricts the invasive capacity of head and neck carcinoma cells harboring a constitutively active PI3 kinase activity
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Brolih, Sanja, Parks, Scott K., Vial, Valérie, Durivault, Jérôme, Mostosi, Livio, Pouysségur, Jacques, Pagès, Gilles, and Picco, Vincent
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- 2018
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18. From gene to mechanics: a comprehensive insight into the mechanobiology of LMNA mutations in cardiomyopathy
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Veltrop, R. J. A., Kukk, M. M., Topouzidou, K., Didden, L., Muchir, A., van Steenbeek, F. G., Schurgers, L. J., and Harakalova, M.
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- 2024
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19. Superbinder based phosphoproteomic landscape revealed PRKCD_pY313 mediates the activation of Src and p38 MAPK to promote TNBC progression
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Deng, Yujiao, Hou, Zhanwu, Li, Yizhen, Yi, Ming, Wu, Ying, Zheng, Yi, Yang, Fei, Zhong, Guansheng, Hao, Qian, Zhai, Zhen, Wang, Meng, Ma, Xiaobin, Kang, Huafeng, Ji, Fanpu, Dong, Chenfang, Liu, Huadong, and Dai, Zhijun
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- 2024
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20. SPOP-mediated ubiquitination and degradation of PDK1 suppresses AKT kinase activity and oncogenic functions
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Yuanzhong Wu, Xiaoling Zhang, Yaqing Su, Xiangpeng Dai, Jianping Guo, Lei Wang, Depei Wu, Xiaomei Zhang, Lang Bu, Wei Xie, Nana Zheng, Xiaomin Zhang, Qiwei Jiang, Shancheng Ren, Wenyi Wei, and Yasheng Zhu
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Cancer Research ,animal structures ,Ubiquitin-Protein Ligases ,P70-S6 Kinase 1 ,SPOP ,Models, Biological ,Cell Line ,3-Phosphoinositide-Dependent Protein Kinases ,Glycogen Synthase Kinase 3 ,Mice ,GSK-3 ,Animals ,Humans ,Phosphorylation ,Protein kinase A ,Protein kinase B ,RC254-282 ,biology ,Kinase ,Research ,AKT ,Ubiquitination ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Nuclear Proteins ,Ubiquitin ligase ,Cell biology ,Repressor Proteins ,Disease Models, Animal ,Cell Transformation, Neoplastic ,Oncology ,PDK1 ,Tumorigenesis ,Mutation ,Proteolysis ,biology.protein ,Molecular Medicine ,Heterografts ,Casein kinase 1 ,CRISPR-Cas Systems ,Proto-Oncogene Proteins c-akt ,Protein Binding - Abstract
Background 3-phosphoinositide-dependent protein kinase-1 (PDK1) acts as a master kinase of protein kinase A, G, and C family (AGC) kinase to predominantly govern cell survival, proliferation, and metabolic homeostasis. Although the regulations to PDK1 downstream substrates such as protein kinase B (AKT) and ribosomal protein S6 kinase beta (S6K) have been well established, the upstream regulators of PDK1, especially its degrader, has not been defined yet. Method A clustered regularly interspaced short palindromic repeats (CRISPR)-based E3 ligase screening approach was employed to identify the E3 ubiquitin ligase for degrading PDK1. Western blotting, immunoprecipitation assays and immunofluorescence (IF) staining were performed to detect the interaction or location of PDK1 with speckle-type POZ protein (SPOP). Immunohistochemistry (IHC) staining was used to study the expression of PDK1 and SPOP in prostate cancer tissues. In vivo and in vitro ubiquitination assays were performed to measure the ubiquitination conjugation of PDK1 by SPOP. In vitro kinase assays and mass spectrometry approach were carried out to identify casein kinase 1 (CK1) and glycogen synthase kinase 3 (GSK3)-mediated PDK1 phosphorylation. The biological effects of PDK1 mutations and correlation with SPOP mutations were performed with colony formation, soft agar assays and in vivo xenograft mouse models. Results We identified that PDK1 underwent SPOP-mediated ubiquitination and subsequent proteasome-dependent degradation. Specifically, SPOP directly bound PDK1 by the consensus degron in a CK1/GSK3β-mediated phosphorylation dependent manner. Pathologically, prostate cancer patients associated mutations of SPOP impaired PDK1 degradation and thus activated the AKT kinase, resulting in tumor malignancies. Meanwhile, mutations that occurred around or within the PDK1 degron, by either blocking SPOP to bind the degron or inhibiting CK1 or GSK3β-mediated PDK1 phosphorylation, could markedly evade SPOP-mediated PDK1 degradation, and played potently oncogenic roles via activating the AKT kinase. Conclusions Our results not only reveal a physiological regulation of PDK1 by E3 ligase SPOP, but also highlight the oncogenic roles of loss-of-function mutations of SPOP or gain-of-function mutations of PDK1 in tumorigenesis through activating the AKT kinase.
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- 2021
21. Inhaled gold nanoparticles cause cerebral edema and upregulate endothelial aquaporin 1 expression, involving caveolin 1 dependent repression of extracellular regulated protein kinase activity
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Yen Ju Chan, Yu Wen Cheng, Chi Hao Tsai, Ching Hao Li, Kuan Hung Lin, Ling Ling Hwang, Ching Yi Chen, Ting Ling Yen, and Po Lin Liao
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MAPK/ERK pathway ,Male ,Gold nanoparticle ,Cell Survival ,Surface Properties ,Health, Toxicology and Mutagenesis ,Caveolin 1 ,lcsh:Industrial hygiene. Industrial welfare ,Metal Nanoparticles ,Brain Edema ,Toxicology ,Cerebral edema ,Cell Line ,Focal adhesion ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Endothelial cell ,lcsh:RA1190-1270 ,medicine ,Animals ,Humans ,Edema ,Particle Size ,Protein kinase A ,Extracellular Signal-Regulated MAP Kinases ,Protein kinase B ,lcsh:Toxicology. Poisons ,030304 developmental biology ,0303 health sciences ,Inhalation Exposure ,Mice, Inbred ICR ,Dose-Response Relationship, Drug ,Aquaporin 1 ,Chemistry ,Research ,Endothelial Cells ,Water ,Correction ,General Medicine ,medicine.disease ,Cell biology ,Endothelial stem cell ,ERK ,030220 oncology & carcinogenesis ,Gold ,lcsh:HD7260-7780.8 - Abstract
Background Gold nanoparticles (Au-NPs) have extensive applications in electronics and biomedicine, resulting in increased exposure and prompting safety concerns for human health. After absorption, nanoparticles enter circulation and effect endothelial cells. We previously showed that exposure to Au-NPs (40–50 nm) collapsed endothelial tight junctions and increased their paracellular permeability. Inhaled nanoparticles have gained significant attention due to their biodistribution in the brain; however, little is known regarding their role in cerebral edema. The present study investigated the expression of aquaporin 1 (AQP1) in the cerebral endothelial cell line, bEnd.3, stimulated by Au-NPs. Results We found that treatment with Au-NPs induced AQP1 expression and increased endothelial permeability to water. Au-NP exposure rapidly boosted the phosphorylation levels of focal adhesion kinase (FAK) and AKT, increased the accumulation of caveolin 1 (Cav1), and reduced the activity of extracellular regulated protein kinases (ERK). The inhibition of AKT (GDC-0068) or FAK (PF-573228) not only rescued ERK activity but also prevented AQP1 induction, whereas Au-NP-mediated Cav1 accumulation remained unaltered. Neither these signaling molecules nor AQP1 expression responded to Au-NPs while Cav1 was silenced. Inhibition of ERK activity (U0126) remarkably enhanced Cav1 and AQP1 expression in bEnd.3 cells. These data demonstrate that Au-NP-mediated AQP1 induction is Cav1 dependent, but requires the repression on ERK activity. Mice receiving intranasally administered Au-NPs displayed cerebral edema, significantly augmented AQP1 protein levels; furthermore, mild focal lesions were observed in the cerebral parenchyma. Conclusions These data suggest that the subacute exposure of nanoparticles might induce cerebral edema, involving the Cav1 dependent accumulation on endothelial AQP1.
- Published
- 2019
22. Cardiovascular function is not associated with creatine kinase activity in a black African population: The SABPA study
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Hugo W. Huisman, Catharina M. C. Mels, and Caitlynd van Zyl
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Male ,Blood Pressure ,030204 cardiovascular system & hematology ,South Africa ,0302 clinical medicine ,Risk Factors ,Ethnicity ,030212 general & internal medicine ,Pulse wave velocity ,education.field_of_study ,biology ,Creatine Kinase, MM Form ,Blood Pressure Monitoring, Ambulatory ,Middle Aged ,Pulse pressure ,Up-Regulation ,medicine.anatomical_structure ,Hypertension ,Female ,Cardiology and Cardiovascular Medicine ,Research Article ,Adult ,medicine.medical_specialty ,Ambulatory blood pressure ,Total peripheral resistance ,Population ,Black People ,Pulse Wave Analysis ,White People ,03 medical and health sciences ,Vascular Stiffness ,Internal medicine ,Creatine kinase activity ,medicine ,Humans ,education ,business.industry ,Health Status Disparities ,Surgery ,Endocrinology ,Blood pressure ,Cross-Sectional Studies ,Vascular resistance ,biology.protein ,Creatine kinase ,Vascular Resistance ,business ,Body mass index ,Biomarkers - Abstract
Background Higher creatine kinase (CK) activity is associated with the development of cardiovascular disease in black African populations. We compared CK activity and investigated associations of blood pressure with CK activity in black and white men as well as black and white women. Methods Ambulatory blood pressure, total peripheral resistance and pulse wave velocity of 197 black and 208 white participants were determined and serum CK activity was measured. Results Blood pressure and pulse wave velocity were higher in black men and women (all p
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- 2016
23. Development of ERK Activity Sensor, an in vitro, FRET-based sensor of Extracellular Regulated Kinase activity
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Green, Harry M., Alberola-Ila, Jose, Green, Harry M., and Alberola-Ila, Jose
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Background: Study of ERK activation has thus far relied on biochemical assays that are limited to the use of phospho-specific antibodies and radioactivity in vitro, and analysis of whole cell populations in vivo. As with many systems, fluorescence resonance energy transfer (FRET) can be utilized to make highly sensitive detectors of molecular activity. Here we introduce FRET-based ERK Activity Sensors, which utilize variants of Enhanced Green Fluorescent Protein fused by an ERK-specific peptide linker to detect ERK2 activity. Results: ERK Activity Sensors display varying changes in FRET upon phosphorylation by active ERK2 in vitro depending on the composition of ERK-specific peptide linker sequences derived from known in vivo ERK targets, Ets1 and Elk1. Analysis of point mutations reveals specific residues involved in ERK binding and phosphorylation of ERK Activity Sensor 3. ERK2 also shows high in vitro specificity for these sensors over two other major MAP Kinases, p38 and pSAPK/JNK. Conclusion: EAS's are a convenient, non-radioactive alternative to study ERK dynamics in vitro. They can be utilized to study ERK activity in real-time. This new technology can be applied to studying ERK kinetics in vitro, analysis of ERK activity in whole cell extracts, and high-throughput screening technologies.
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- 2005
24. Alantolactone, a natural sesquiterpene lactone, has potent antitumor activity against glioblastoma by targeting IKKß kinase activity and interrupting NF-κB/COX-2-mediated signaling cascades.
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Xun Wang, Zhenlong Yu, Chao Wang, Wei Cheng, Xiangge Tian, Xiaokui Huo, Yan Wang, Chengpeng Sun, Lei Feng, Jinshan Xing, Yulong Lan, Dongdong Sun, Qingjuan Hou, Baojing Zhang, Xiaochi Ma, and Bo Zhang
- Subjects
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SESQUITERPENE lactones , *ANTINEOPLASTIC agents , *GLIOBLASTOMA multiforme treatment , *KINASES , *NF-kappa B , *CYCLOOXYGENASE 2 , *APOPTOSIS , *CENTRAL nervous system cancer - Abstract
Background: Glioblastoma multiforme (GBM) is one of the most refractory and palindromic central nervous system (CNS) neoplasms, and current treatments have poor effects in GBM patients. Hence, the identification of novel therapeutic targets and the development of effective treatment strategies are essential. Alantolactone (ATL) has a wide range of pharmacological activities, and its anti-tumor effect is receiving increasing attention. However, the molecular mechanism underlying the anti-GBM activity of ATL remains poorly understood. Methods: The biological functions of ATL in GBM cells were investigated using migration/invasion, colony formation and cell cycle/apoptosis assays. The localization of nuclear factor kappa B (NF-κB) p50/p65 and its binding to the cyclooxygenase 2 (COX-2) promoter were determined using confocal immunofluorescence, a streptavidin-agarose pulldown assay and a chromatin immunoprecipitation (ChIP) assay. IKKß kinase activity was determined using a cell IKKß kinase activity spectrophotometry quantitative detection kit and a molecular docking study. LC-MS/MS analysis was performed to determine the ability of ATL to traverse the blood-brain barrier (BBB). The in vivo anti-tumor efficacy of ATL was also analyzed in xenografted nude mice. Western blot analysis was performed to detect the protein expression levels. Results: ATL significantly suppressed the growth of GBM in vivo and in vitro. ATL significantly reduced the expression of COX-2 by inhibiting the kinase activity of IKKß by targeting the ATP-binding site and then attenuating the binding of NF-κB to the COX-2 promoter region. Furthermore, ATL induced apoptosis by activating the cytochrome c (cyt c)/caspase cascade signaling pathway. Moreover, ATL could penetrate the BBB. Conclusions: ATL exerts its anti-tumor effects in human GBM cells at least in part via NF-κB/COX-2-mediated signaling cascades by inhibiting IKKß kinase activity. ATL, which is a natural small molecule inhibitor, is a promising candidate for clinical applications in the treatment of CNS tumors. [ABSTRACT FROM AUTHOR]
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- 2017
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25. The PINK1 p.I368N mutation affects protein stability and ubiquitin kinase activity.
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Ando, Maya, Fiesel, Fabienne C., Hudec, Roman, Caulfield, Thomas R., Kotaro Ogaki, Górka-Skoczylas, Paulina, Koziorowski, Dariusz, Friedman, Andrzej, Li Chen, Dawson, Valina L., Dawson, Ted M., Guojun Bu, Ross, Owen A., Wszolek, Zbigniew K., and Springer, Wolfdieter
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PARKINSON'S disease , *UBIQUITIN , *AUTOPHAGY , *MITOCHONDRIA , *UBIQUITIN ligases - Abstract
Background: Mutations in PINK1 and PARKIN are the most common causes of recessive early-onset Parkinson's disease (EOPD). Together, the mitochondrial ubiquitin (Ub) kinase PINK1 and the cytosolic E3 Ub ligase PARKIN direct a complex regulated, sequential mitochondrial quality control. Thereby, damaged mitochondria are identified and targeted to degradation in order to prevent their accumulation and eventually cell death. Homozygous or compound heterozygous loss of either gene function disrupts this protective pathway, though at different steps and by distinct mechanisms. While structure and function of PARKIN variants have been well studied, PINK1 mutations remain poorly characterized, in particular under endogenous conditions. A better understanding of the exact molecular pathogenic mechanisms underlying the pathogenicity is crucial for rational drug design in the future. Methods: Here, we characterized the pathogenicity of the PINK1 p.I368N mutation on the clinical and genetic as well as on the structural and functional level in patients' fibroblasts and in cell-based, biochemical assays. Results: Under endogenous conditions, PINK1 p.I368N is expressed, imported, and N-terminally processed in healthy mitochondria similar to PINK1 wild type (WT). Upon mitochondrial damage, however, full-length PINK1 p.I368N is not sufficiently stabilized on the outer mitochondrial membrane (OMM) resulting in loss of mitochondrial quality control. We found that binding of PINK1 p.I368N to the co-chaperone complex HSP90/CDC37 is reduced and stress-induced interaction with TOM40 of the mitochondrial protein import machinery is abolished. Analysis of a structural PINK1 p.I368N model additionally suggested impairments of Ub kinase activity as the ATP-binding pocket was found deformed and the substrate Ub was slightly misaligned within the active site of the kinase. Functional assays confirmed the lack of Ub kinase activity. Conclusions: Here we demonstrated that mutant PINK1 p.I368N can not be stabilized on the OMM upon mitochondrial stress and due to conformational changes in the active site does not exert kinase activity towards Ub. In patients' fibroblasts, biochemical assays and by structural analyses, we unraveled two pathomechanisms that lead to loss of function upon mutation of p.I368N and highlight potential strategies for future drug development. [ABSTRACT FROM AUTHOR]
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- 2017
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26. Erratum to: Kinase Activity Profiling of Gram-Negative Pneumonia
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Hoogendijk, Arie J, Diks, Sander H, Peppelenbosch, Maikel P, van der Poll, Tom, and Wieland, Catharina W
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- 2011
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27. Dynamic single cell measurements of kinase activity by synthetic kinase activity relocation sensors.
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Durandau, Eric, Aymoz, Delphine, and Pelet, Serge
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MITOGEN-activated protein kinases , *EXTRACELLULAR signal-regulated kinases , *BIOSENSORS , *CELL cycle , *FLUORESCENCE microscopy - Abstract
Background: Mitogen activated protein kinases (MAPK) play an essential role in integrating extra-cellular signals and intra-cellular cues to allow cells to grow, adapt to stresses, or undergo apoptosis. Budding yeast serves as a powerful system to understand the fundamental regulatory mechanisms that allow these pathways to combine multiple signals and deliver an appropriate response. To fully comprehend the variability and dynamics of these signaling cascades, dynamic and quantitative single cell measurements are required. Microscopy is an ideal technique to obtain these data; however, novel assays have to be developed to measure the activity of these cascades. Results: We have generated fluorescent biosensors that allow the real-time measurement of kinase activity at the single cell level. Here, synthetic MAPK substrates were engineered to undergo nuclear-to-cytoplasmic relocation upon phosphorylation of a nuclear localization sequence. Combination of fluorescence microscopy and automated image analysis allows the quantification of the dynamics of kinase activity in hundreds of single cells. A large heterogeneity in the dynamics of MAPK activity between individual cells was measured. The variability in the mating pathway can be accounted for by differences in cell cycle stage, while, in the cell wall integrity pathway, the response to cell wall stress is independent of cell cycle stage. Conclusions: These synthetic kinase activity relocation sensors allow the quantification of kinase activity in live single cells. The modularity of the architecture of these reporters will allow their application in many other signaling cascades. These measurements will allow to uncover new dynamic behaviour that previously could not be observed in population level measurements. [ABSTRACT FROM AUTHOR]
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- 2015
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28. Alantolactone, a natural sesquiterpene lactone, has potent antitumor activity against glioblastoma by targeting IKKβ kinase activity and interrupting NF-κB/COX-2-mediated signaling cascades.
- Author
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Wang X, Yu Z, Wang C, Cheng W, Tian X, Huo X, Wang Y, Sun C, Feng L, Xing J, Lan Y, Sun D, Hou Q, Zhang B, Ma X, and Zhang B
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- Adenosine Triphosphate chemistry, Adenosine Triphosphate metabolism, Animals, Antineoplastic Agents chemistry, Apoptosis drug effects, Binding Sites, Biomarkers, Blood-Brain Barrier metabolism, Caspases metabolism, Cell Cycle Checkpoints drug effects, Cell Line, Tumor, Cell Proliferation drug effects, Cell Survival drug effects, Cytochromes c metabolism, Disease Models, Animal, Dose-Response Relationship, Drug, E1A-Associated p300 Protein metabolism, Humans, I-kappa B Kinase chemistry, Lactones chemistry, Male, Mice, Models, Biological, Molecular Conformation, Promoter Regions, Genetic, Protein Binding, Rats, Sesquiterpenes, Eudesmane chemistry, Antineoplastic Agents pharmacology, Cyclooxygenase 2 metabolism, I-kappa B Kinase antagonists & inhibitors, Lactones pharmacology, NF-kappa B metabolism, Sesquiterpenes, Eudesmane pharmacology, Signal Transduction drug effects
- Abstract
Background: Glioblastoma multiforme (GBM) is one of the most refractory and palindromic central nervous system (CNS) neoplasms, and current treatments have poor effects in GBM patients. Hence, the identification of novel therapeutic targets and the development of effective treatment strategies are essential. Alantolactone (ATL) has a wide range of pharmacological activities, and its anti-tumor effect is receiving increasing attention. However, the molecular mechanism underlying the anti-GBM activity of ATL remains poorly understood., Methods: The biological functions of ATL in GBM cells were investigated using migration/invasion, colony formation and cell cycle/apoptosis assays. The localization of nuclear factor kappa B (NF-κB) p50/p65 and its binding to the cyclooxygenase 2 (COX-2) promoter were determined using confocal immunofluorescence, a streptavidin-agarose pulldown assay and a chromatin immunoprecipitation (ChIP) assay. IKKβ kinase activity was determined using a cell IKKβ kinase activity spectrophotometry quantitative detection kit and a molecular docking study. LC-MS/MS analysis was performed to determine the ability of ATL to traverse the blood-brain barrier (BBB). The in vivo anti-tumor efficacy of ATL was also analyzed in xenografted nude mice. Western blot analysis was performed to detect the protein expression levels., Results: ATL significantly suppressed the growth of GBM in vivo and in vitro. ATL significantly reduced the expression of COX-2 by inhibiting the kinase activity of IKKβ by targeting the ATP-binding site and then attenuating the binding of NF-κB to the COX-2 promoter region. Furthermore, ATL induced apoptosis by activating the cytochrome c (cyt c)/caspase cascade signaling pathway. Moreover, ATL could penetrate the BBB., Conclusions: ATL exerts its anti-tumor effects in human GBM cells at least in part via NF-κB/COX-2-mediated signaling cascades by inhibiting IKKβ kinase activity. ATL, which is a natural small molecule inhibitor, is a promising candidate for clinical applications in the treatment of CNS tumors.
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- 2017
- Full Text
- View/download PDF
29. TGFβ1-induced SMAD2/3 and SMAD1/5 phosphorylation are both ALK5-kinase-dependent in primary chondrocytes and mediated by TAK1 kinase activity.
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van Caam A, Madej W, Garcia de Vinuesa A, Goumans MJ, Ten Dijke P, Blaney Davidson E, and van der Kraan P
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- Animals, Benzodioxoles pharmacology, Cattle, Chondrocytes drug effects, Imidazoles pharmacology, Phosphorylation, Pyrazoles pharmacology, Pyridines pharmacology, Pyrimidines pharmacology, Receptor, Transforming Growth Factor-beta Type I, Transforming Growth Factor beta1 metabolism, Zearalenone analogs & derivatives, Zearalenone pharmacology, Chondrocytes metabolism, Enzyme Inhibitors pharmacology, MAP Kinase Kinase Kinases metabolism, Osteoarthritis metabolism, Protein Serine-Threonine Kinases metabolism, Receptors, Transforming Growth Factor beta metabolism, Smad Proteins metabolism
- Abstract
Background: Dysregulated transforming growth factor β (TGFβ) signaling is implicated in osteoarthritis development, making normalizing TGFβ signaling a possible therapy. Theoretically, this can be achieved with small molecule inhibitors specifically targeting the various TGFβ receptors and downstream mediators. In this study we explore in primary chondrocytes the use of small molecule inhibitors to target TGFβ-induced pSmad1/5/9-, pSmad2/3- and TGFβ-activated kinase 1 (TAK1)-dependent signaling., Method: Primary bovine chondrocytes and explants were isolated from metacarpophalangeal joints. To modulate TGFβ signaling the activin receptor-like kinase (ALK)1/2/3/6 inhibitor LDN-193189, the ALK4/5/7 inhibitor SB-505124 and the TAK1 inhibitor (5Z)-7-Oxozeaenol were used. pSmad1/5 and pSmad2 were measured using western blot analysis and TGFβ1-induced gene expression was measured using quantitative real time PCR (qPCR)., Results: In chondrocytes, TGFβ1 strongly induced both pSmad1/5 and pSmad2. Remarkably, LDN-193189 did not inhibit TGFβ-induced pSmad1/5. In contrast, SB-505124 did inhibit both TGFβ-induced Smad2 and Smad1/5 phosphorylation. Furthermore, (5Z)-7-Oxozeaenol also profoundly inhibited TGFβ-induced pSmad2 and pSmad1/5. Importantly, both SB-505124 and (5Z)-7-Oxozeaenol did not significantly inhibit constitutively active ALK1, making an off-target effect unlikely. Additionally, LDN-193189 was able to potently inhibit BMP2/7/9-induced pSmad1/5, showing its functionality. On gene expression, LDN-193189 did not affect TGFβ1-induced regulation, whereas both SB-505124 and (5Z)-7-Oxozeaenol did. Similar results were obtained in cartilage explants, although pSmad1/5 was not strongly induced by addition of TGFβ1., Conclusion: Our data suggest that ALK5 kinase activity plays a central role in both TGFβ-induced Smad1/5 and Smad2/3 phosphorylation, making it difficult to separate both pathways with the use of currently available small molecule inhibitors. Furthermore, our data regarding (5Z)-7-Oxozeaenol suggest that TAK1 facilitates Smad-dependent signaling.
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- 2017
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30. Cardiovascular function is not associated with creatine kinase activity in a black African population: The SABPA study.
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Mels CM, van Zyl C, and Huisman HW
- Subjects
- Adult, Biomarkers blood, Blood Pressure Monitoring, Ambulatory, Cross-Sectional Studies, Female, Humans, Hypertension blood, Hypertension physiopathology, Male, Middle Aged, Pulse Wave Analysis, Risk Factors, South Africa, Up-Regulation, Vascular Resistance, Vascular Stiffness, Black People, Blood Pressure, Creatine Kinase, MM Form blood, Health Status Disparities, Hypertension diagnosis, Hypertension ethnology, White People
- Abstract
Background: Higher creatine kinase (CK) activity is associated with the development of cardiovascular disease in black African populations. We compared CK activity and investigated associations of blood pressure with CK activity in black and white men as well as black and white women., Methods: Ambulatory blood pressure, total peripheral resistance and pulse wave velocity of 197 black and 208 white participants were determined and serum CK activity was measured., Results: Blood pressure and pulse wave velocity were higher in black men and women (all p < 0.001) when compared to their white counterparts. CK activity only varied between black and white women (75.9 U/l vs 62.8 U/l, p = 0.009), even after adjusting for age, body mass index and physical activity. Despite the worse cardiovascular profile of black men and women, and the higher CK activity in the black women, we were unable to link blood pressure, pulse wave velocity or total peripheral resistance with CK activity, in the black African population. In white men, total peripheral resistance was associated with CK activity (R (2) = 0.32; β = 0.25; p = 0.009), whereas systolic blood pressure (R (2) = 0.46; β = 0.17; p = 0.03) and pulse pressure (R (2) = 0.31; β = 0.21; p = 0.01) were associated with CK activity in white women., Conclusions: The lack of associations in the black African population suggests that the link between a worse cardiovascular profile and CK activity may be overshadowed by other contributing factors. Whereas, the established link between cardiovascular function and CK activity in the white groups may be the result of enhanced smooth muscle cell contractility and/or attenuated nitric oxide synthesis capacity.
- Published
- 2016
- Full Text
- View/download PDF
31. Development of ERK Activity Sensor, an in vitro, FRET-based sensor of Extracellular Regulated Kinase activity
- Author
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Harry M Green and José Alberola-Ila
- Subjects
MAPK/ERK pathway ,Kinase ,p38 mitogen-activated protein kinases ,Biology ,Cell biology ,lcsh:Biochemistry ,Förster resonance energy transfer ,Biochemistry ,ELK1 ,In vivo ,Drug Discovery ,Molecular Medicine ,Phosphorylation ,lcsh:QD415-436 ,Kinase activity ,Caltech Library Services ,Research Article - Abstract
Background Study of ERK activation has thus far relied on biochemical assays that are limited to the use of phospho-specific antibodies and radioactivity in vitro, and analysis of whole cell populations in vivo. As with many systems, fluorescence resonance energy transfer (FRET) can be utilized to make highly sensitive detectors of molecular activity. Here we introduce FRET-based ERK Activity Sensors, which utilize variants of Enhanced Green Fluorescent Protein fused by an ERK-specific peptide linker to detect ERK2 activity. Results ERK Activity Sensors display varying changes in FRET upon phosphorylation by active ERK2 in vitro depending on the composition of ERK-specific peptide linker sequences derived from known in vivo ERK targets, Ets1 and Elk1. Analysis of point mutations reveals specific residues involved in ERK binding and phosphorylation of ERK Activity Sensor 3. ERK2 also shows high in vitro specificity for these sensors over two other major MAP Kinases, p38 and pSAPK/JNK. Conclusion EAS's are a convenient, non-radioactive alternative to study ERK dynamics in vitro. They can be utilized to study ERK activity in real-time. This new technology can be applied to studying ERK kinetics in vitro, analysis of ERK activity in whole cell extracts, and high-throughput screening technologies.
- Published
- 2005
32. H2AX phosphorylation and DNA damage kinase activity are dispensable for herpes simplex virus replication.
- Author
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Botting, Carolyn, Xu Lu, and Triezenberg, Steven J.
- Subjects
- *
PHOSPHORYLATION , *CHEMICAL reactions , *DNA damage , *BIOCHEMICAL genetics , *HERPES simplex virus - Abstract
Background: Herpes simplex virus type 1 (HSV-1) can establish both lytic and latent infections in humans. The phosphorylation of histone H2AX, a common marker of DNA damage, during lytic infection by HSV-1 is well established. However, the role(s) of H2AX phosphorylation in lytic infection remain unclear. Methods: Following infection of human foreskin fibroblasts by HSV-1 or HSV-2, we assayed the phosphorylation of H2AX in the presence of inhibitors of transcription, translation, or viral DNA replication, or in the presence of inhibitors of ATM and ATR kinases (KU-55933 and VE-821, respectively). We also assayed viral replication in fibroblasts in the presence of the kinase inhibitors or siRNAs specific for ATM and ATR, as well as in cell lines deficient for either ATR or ATM. Results: The expression of viral immediate-early and early proteins (including the viral DNA polymerase), but not viral DNA replication or late protein expression, were required for H2AX phosphorylation following HSV-1 infection. Inhibition of ATM kinase activity prevented HSV-stimulated H2AX phosphorylation but had only a minor effect on DNA replication and virus yield in HFF cells. These results differ from previous reports of a dramatic reduction in viral yield following chemical inhibition of ATM in oral keratinocytes or following infection of ATM-/- cells. Inhibition of the closely related kinase ATR (whether by chemical inhibitor or siRNA disruption) had no effect on H2AX phosphorylation and reduced viral DNA replication only moderately. During infection by HSV-2, H2AX phosphorylation was similarly dispensable but was dependent on both ATM activity and viral DNA replication. Conclusion: H2AX phosphorylation represents a cell type-specific and virus type-specific host response to HSV infection with little impact on viral infection. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
33. Metformin enhances nitric oxide production and diminishes Rho kinase activity in rats with hyperlipidemia.
- Author
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Yan Liu, Congwu Huang, Chuan Ceng, Haiyong Zhan, Dongdan Zheng, and Weixing Han
- Abstract
Background: Rho kinase over-activation is associated with nitric oxide (NO) reduction and atherosclerosis. Metformin is favorable for endothelial function improvement and cardiovascular outcomes. Whether cardio-protective effect of metformin is associated with Rho kinase activity is unknown. Methods: Hyperlipidemia model of rats were established accordingly. Thereafter, medical interventions in terms of atorvastatin, metformin or combined therapy were administered for 4 weeks. Laboratory parameters were compared among each groups at initial, 6 weeks of high-fat and high-cholesterol diet administration, and 4 weeks of medical intervention. Lineal regression analyses were performed. Results: No significant difference of laboratory parameters was observed initially. Six weeks of high-fat and high-cholesterol diet administration, serum levels of cholesterol, C-reactive protein (CRP) level, and Rho kinase activity were significantly increased while NO production was concomitantly reduced in comparison to the sham group. After 4 weeks of medical intervention, CRP level and Rho kinase activity were profoundly diminished while NO production was significantly enhanced in the atorvastatin and metformin groups, and these benefits were further enhanced with combined therapy. Lineal regression analyses showed that Rho kinase activity was negatively correlated with NO production but positively correlated with CRP level. Conclusion: In rats with hyperlipidemia, metformin and atorvastatin therapy is favorable for NO production and CRP reduction, which might be associated with Rho kinase activity decrease. [ABSTRACT FROM AUTHOR]
- Published
- 2014
- Full Text
- View/download PDF
34. Is inhibition of kinase activity the only therapeutic strategy for LRRK2-associated Parkinson's disease?
- Subjects
- *
KINASE inhibitors , *PROTEIN kinases , *GENETIC mutation , *PARKINSON'S disease , *PROTEIN binding , *THERAPEUTICS - Abstract
The article presents a study whether inhibition of kinase activity is the only therapeutic strategy for LRRK2-associated Parkinson's disease. LRRK2 has a complex multidomain structure. The central part of the protein contains three independent domains. LRRK2 is an active protein kinase, although the true physiological target of this kinase activity has not been resolved
- Published
- 2012
- Full Text
- View/download PDF
35. Kinase Activity Profiling of Gram-Negative Pneumonia.
- Author
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Hoogendijk, Arie J., Diks, Sander H., Peppelenbosch, Maikel P., der Poll, Tom van, and Wieland, Catharina W.
- Subjects
- *
PNEUMONIA , *MITOGEN-activated protein kinases , *ONCOGENES , *GLYCOGEN synthase kinase-3 , *KLEBSIELLA pneumoniae , *PATTERN perception , *PEPTIDOGLYCANS , *PHOSPHORYLATION - Abstract
Pneumonia is a severe disease with high morbidity and mortality. A major causative pathogen is the Gram-negative bacterium Klebsiella (K.) pneumoniae. Kinases play an integral role in the transduction of intracellular signaling cascades and regulate a diverse array of biological processes essential to immune cells. The current study explored signal transduction events during murine Gram-negative pneumonia using a systems biology approach. Kinase activity arrays enable the analysis of 1,024 consensus sequences of protein kinase substrates. Using a kinase activity array on whole lung lysates, cellular kinase activities were determined in a mouse model of K. pneumoniae pneumonia. Notable kinase activities also were validated with phospho-specific Western blots. On the basis of the profiling data, mitogen-activated protein kinase (MAPK) signaling via p42 mitogen-activated protein kinase (p42) and p38 mitogen-activated protein kinase (p38) and transforming growth factor β (TGFβ) activity were reduced during infection, whereas v-src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog (avian) (SRC) activity generally was enhanced. AKT signaling was represented in both metabolic and inflammatory (mitogen-activated protein kinase kinase 2 [MKK], apoptosis signal-regulating kinase/mitogen-activated protein kinase kinase kinase 5 [ASK] and v-raf murine sarcoma viral oncogene homolog B1 [b-RAF]) context. This study reaffirms the importance of classic inflammation pathways, such as MAPK and TGFβ signaling and reveals less known involvement of glycogen synthase kinase 3β (GSK-3β), AKT and SRC signaling cassettes in pneumonia. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
36. Phosphoinositide-3 kinase gamma kinase activity significantly contributes to the pathophysiology of sepsis and multiorgan failure
- Author
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V. Marco Ranieri, Luisa Delsedime, Emilio Hirsch, E. L. Martin, and Martino Bosco
- Subjects
Pathology ,medicine.medical_specialty ,Phosphoinositide 3-kinase ,biology ,business.industry ,Critical Care and Intensive Care Medicine ,Bioinformatics ,medicine.disease ,Multiorgan failure ,Pathophysiology ,Sepsis ,Poster Presentation ,biology.protein ,Medicine ,Kinase activity ,business - Published
- 2008
37. Dispensable role of Drosophila ortholog of LRRK2 kinase activity in survival of dopaminergic neurons.
- Author
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Danling Wang, Beisha Tang, Guohua Zhao, Qian Pan, Kun Xia, Bodmer, Rolf, and Zhuohua Zhang
- Subjects
- *
GENES , *PROTEIN kinases , *DROSOPHILA genetics , *DOPAMINERGIC neurons , *PARKINSON'S disease & genetics , *NEURODEGENERATION , *GENETIC mutation , *OXIDATIVE stress - Abstract
Background: Parkinson's disease (PD) is the most prevalent incurable neurodegenerative movement disorder. Mutations in LRRK2 are associated with both autosomal dominant familial and sporadic forms of PD. LRRK2 encodes a large putative serine/threonine kinase with GTPase activity. Increased LRRK2 kinase activity plays a critical role in pathogenic LRRK2 mutant-induced neurodegeneration in vitro. Little is known about the physiological function of LRRK2. Results: We have recently identified a Drosophila line with a P-element insertion in an ortholog gene of human LRRK2 (dLRRK). The insertion results in a truncated Drosophila LRRK variant with N-terminal 1290 amino acids but lacking C-terminal kinase domain. The homozygous mutant fly develops normally with normal life span as well as unchanged number and pattern of dopaminergic neurons. However, dLRRK mutant flies were selectively sensitive to hydrogen peroxide induced stress but not to paraquat, rotenone and β-mercaptoethanol induced stresses. Conclusion: Our results indicate that inactivation of dLRRK kinase activity is not essential for fly development and suggest that inhibition of LRRK activity may serve as a potential treatment of PD. However, dLRRK kinase activity likely plays a role in protecting against oxidative stress. [ABSTRACT FROM AUTHOR]
- Published
- 2008
- Full Text
- View/download PDF
38. Metformin enhances nitric oxide production and diminishes Rho kinase activity in rats with hyperlipidemia.
- Author
-
Liu Y, Huang C, Ceng C, Zhan H, Zheng D, and Han W
- Subjects
- Animals, Anticholesteremic Agents therapeutic use, Atorvastatin, C-Reactive Protein metabolism, Drug Evaluation, Preclinical, Heptanoic Acids therapeutic use, Hypercholesterolemia blood, Hypercholesterolemia enzymology, Male, Metformin therapeutic use, Nitric Oxide blood, Pyrroles therapeutic use, Rats, Sprague-Dawley, Anticholesteremic Agents pharmacology, Heptanoic Acids pharmacology, Hypercholesterolemia drug therapy, Metformin pharmacology, Nitric Oxide biosynthesis, Pyrroles pharmacology, rho-Associated Kinases blood
- Abstract
Background: Rho kinase over-activation is associated with nitric oxide (NO) reduction and atherosclerosis. Metformin is favorable for endothelial function improvement and cardiovascular outcomes. Whether cardio-protective effect of metformin is associated with Rho kinase activity is unknown., Methods: Hyperlipidemia model of rats were established accordingly. Thereafter, medical interventions in terms of atorvastatin, metformin or combined therapy were administered for 4 weeks. Laboratory parameters were compared among each groups at initial, 6 weeks of high-fat and high-cholesterol diet administration, and 4 weeks of medical intervention. Lineal regression analyses were performed., Results: No significant difference of laboratory parameters was observed initially. Six weeks of high-fat and high-cholesterol diet administration, serum levels of cholesterol, C-reactive protein (CRP) level, and Rho kinase activity were significantly increased while NO production was concomitantly reduced in comparison to the sham group. After 4 weeks of medical intervention, CRP level and Rho kinase activity were profoundly diminished while NO production was significantly enhanced in the atorvastatin and metformin groups, and these benefits were further enhanced with combined therapy. Lineal regression analyses showed that Rho kinase activity was negatively correlated with NO production but positively correlated with CRP level., Conclusion: In rats with hyperlipidemia, metformin and atorvastatin therapy is favorable for NO production and CRP reduction, which might be associated with Rho kinase activity decrease.
- Published
- 2014
- Full Text
- View/download PDF
39. ATP directed agent, 8-chloro-adenosine, induces AMP activated protein kinase activity, leading to autophagic cell death in breast cancer cells.
- Author
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Stellrecht CM, Vangapandu HV, Le XF, Mao W, and Shentu S
- Subjects
- 2-Chloroadenosine pharmacology, Animals, Autophagy drug effects, Breast Neoplasms metabolism, Breast Neoplasms pathology, Cell Line, Tumor, Disease Models, Animal, Female, Humans, MCF-7 Cells, Mice, Phosphorylation drug effects, Random Allocation, Signal Transduction drug effects, Xenograft Model Antitumor Assays, 2-Chloroadenosine analogs & derivatives, AMP-Activated Protein Kinases metabolism, Apoptosis drug effects, Breast Neoplasms drug therapy
- Abstract
Background: 8-chloro-adenosine (8-Cl-Ado) is a unique ribonucleoside analog which is currently in a phase I clinical trial for hematological malignancies. Previously, we demonstrated in breast cancer cells that a 3-day treatment with 10 μM 8-Cl-Ado causes a 90% loss of clonogenic survival. In contrast, there was only a modest induction of apoptosis under these conditions, suggesting an alternative mechanism for the tumoricidal activity of 8-Cl-Ado., Methods: Cellular metabolism, AMP-activated protein kinase (AMPK) and mammalian target of rapamycin (mTOR) pathway signaling, as well as autophagy induction was evaluated in breast cancer cell lines treated with 8-Cl-Ado. The effects of knocking down essential autophagy factors with small interfering RNA on 8-Cl-Ado-inhibited cell survival was assessed in breast cancer cells by examining apoptosis induction and clonogenic survival. In vivo efficacy of 8-Cl-Ado was measured in two breast cancer orthotopic model systems., Results: We demonstrate that in breast cancer cell lines, the metabolism of 8-Cl-Ado results in depletion of endogenous ATP that subsequently induces the phosphorylation and activation of the energy sensor, AMPK. This was associated with an attenuation of mTOR signaling and an induction of the phosphorylation of the autophagy factor, Unc51-like kinase 1 on Ser555. 8-Cl-Ado-mediated induction of autophagy was evident by increased aggregates of microtubule-associated protein 1 light chain 3B (LC3B) which was associated with its conversion to its lipidated form, LC3B-II, p62 degradative flux, and increased formation of acidic vesicular organelles. Additionally, transfection of MCF-7 cells with siRNA to ATG7 or beclin 1 provided partial protection of the cells to 8-Cl-Ado cytotoxicity as measured by clonogenicity. In vivo, 8-Cl-Ado inhibited growth of both MCF-7 and BT-474 xenograft tumors. Moreover, in 9 of 22 BT-474 tumors treated with 100 mg/kg/day 3 times a week, there was an absence of macroscopically detectable tumor after 3 weeks of treatment., Conclusions: Our data demonstrates that 8-Cl-Ado treatment activates the AMPK pathway leading to autophagy induction of in breast cancer cells, eliciting, in part, its tumoricidal effects. Additionally, 8-Cl-Ado effectively inhibited in vivo tumor growth in mice. Based on this biological activity, we are planning to test 8-Cl-Ado in the clinic for patients with breast cancer.
- Published
- 2014
- Full Text
- View/download PDF
40. Protein kinase activity is associated with CD63 in melanoma cells.
- Author
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Iida, Joji, Skubitz, Amy P. N., McCarthy, James B., and Skubitz, Keith M.
- Subjects
- *
PROTEIN kinases , *MELANOMA , *CANCER cells , *ANTIGENS , *EXTRACELLULAR matrix proteins , *SERUM - Abstract
Background: The tetraspan protein CD63, originally described as a stage-specific melanoma antigen but also present in a number of normal cells, regulates melanoma cell growth in nude mice, motility in serum containing media, and adhesion to several extracellular matrix proteins. CD63 has been reported to associate with β1 and β2 integrins, but the mechanism of signal transduction by CD63 is not clear. This study examined whether CD63 is associated with protein kinase and can transmit signals in melanoma cells. Methods: Immunoprecipitation and radiolabeling were used to test for association of protein kinase activity with CD63. Adhesion of cells to monoclonal antibodies immobilized to microtiter plates was used to examine the ability of CD63 to transmit signals. Results: CD63 was capable of transmitting a signal in melanoma cells that required extracellular calcium. In the absence of extracellular calcium at the time of binding to the CD63 mAb, the cell was no longer responsive to stimulation by CD63. Immunoprecipitation studies demonstrated protein kinase activity associated with CD63, and phosphoamino acid analysis revealed that most of this protein kinase activity was due to serine kinase activity. Conclusion: The current study suggests that serine protein kinase activity associated with CD63 may play a role in signaling by CD63 in melanoma cells. [ABSTRACT FROM AUTHOR]
- Published
- 2005
- Full Text
- View/download PDF
41. Kinotypes: stable species- and individual-specific profiles of cellular kinase activity.
- Author
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Trost B, Kindrachuk J, Scruten E, Griebel P, Kusalik A, and Napper S
- Subjects
- Adult, Animals, Cluster Analysis, Enzyme Activation, Female, Gene Regulatory Networks, Humans, Male, Middle Aged, Peptides genetics, Peptides metabolism, Phosphotransferases genetics, Protein Interaction Mapping, Protein Interaction Maps, Species Specificity, Swine, Young Adult, Leukocytes, Mononuclear enzymology, Phosphotransferases metabolism, Proteome, Proteomics
- Abstract
Background: Recently, questions have been raised regarding the ability of animal models to recapitulate human disease at the molecular level. It has also been demonstrated that cellular kinases, individually or as a collective unit (the kinome), play critical roles in regulating complex biology. Despite the intimate relationship between kinases and health, little is known about the variability, consistency and stability of kinome profiles across species and individuals., Results: As a preliminary investigation of the existence of species- and individual-specific kinotypes (kinome signatures), peptide arrays were employed for the analysis of peripheral blood mononuclear cells collected weekly from human and porcine subjects (n = 6) over a one month period. The data revealed strong evidence for species-specific signalling profiles. Both humans and pigs also exhibited evidence for individual-specific kinome profiles that were independent of natural changes in blood cell populations., Conclusions: Species-specific kinotypes could have applications in disease research by facilitating the selection of appropriate animal models or by revealing a baseline kinomic signature to which treatment-induced profiles could be compared. Similarly, individual-specific kinotypes could have implications in personalized medicine, where the identification of molecular patterns or signatures within the kinome may depend on both the levels of kinome diversity and temporal stability across individuals.
- Published
- 2013
- Full Text
- View/download PDF
42. SU11652 Inhibits tyrosine kinase activity of FLT3 and growth of MV-4-11 cells.
- Author
-
Yao Guo, Yun Chen, Xuesong Xu, Xueqi Fu, and Zhizhuang Joe Zhao
- Subjects
- *
PROTEIN-tyrosine kinases , *MYELOID leukemia , *DRUG development , *APOPTOSIS , *CELL cycle , *ENZYME inhibitors - Abstract
Background: FLT3-ITD and FLT3-TKD mutations are frequently found in acute myeloid leukemia (AML). This makes tyrosine kinase FLT3 a highly attractive target for therapeutic drug development. However, effective drugs have not yet emerged. This study is intended to identify and to characterize new FLT3 inhibitors. Methods: By using the protein substrate GST-FLT3S to analyze kinase activity of recombinant proteins carrying the catalytic domain of wild type and mutant forms of FLT3, we screened a chemical library containing 80 known protein kinase inhibitors. We identified SU11652 as a potent FLT3 inhibitor and further employed FLT3-ITD-positive MV- 4-11 cells to study its effects on cell growth, apoptosis, cell cycles, and cell signaling. Results: SU11652 strongly inhibited the activity of wild type, D835Y, and D835H mutant forms of FLT3 with IC50 values of 1.5, 16, and 32 nM, respectively. It effectively blocked the growth of FLT3-ITD -positive MV-4-11 cells at nanomolar concentrations but exhibited much less effects on several other cells which do not carry mutations of FLT3. SU11652 inhibited growth of MV-4-11 cells by inducing apoptosis, causing cell cycle arrest, and blocking activation of the ERK, Akt, and STAT signaling pathways. Conclusion: SU11652 is a potent FLT3 inhibitor which selectively targets FLT3-ITD-positive cells. It should serve as a good candidate for development of therapeutic drugs to treat AML [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
43. Inhibition of Src kinase activity attenuates amyloid associated microgliosis in a murine model of Alzheimer's disease.
- Subjects
- *
SRC gene , *AMYLOID beta-protein , *PROTEIN-tyrosine kinases , *ALZHEIMER'S disease , *MICROGLIA , *NEUROLOGY - Abstract
The article offers information on a research that studies the inhibition of amyloid beta peptide stimulation of microglia as therapeutic intervention for Alzheimer's disease (AD). It is mentioned that activation of microglia is an important pathological finding of the AD and microglia is believed to be involved in pro-inflammatory changes associated with the disease. Also, microgliosis in the diseased brain is found to be dependent upon the Src kinase activity.
- Published
- 2012
- Full Text
- View/download PDF
44. Autophosphorylation of serine 608 in the p85 regulatory subunit of wild type or cancer-associated mutants of phosphoinositide 3-kinase does not affect its lipid kinase activity.
- Author
-
Layton, Meredith J., Saad, Mirette, Church, Nicole L., Pearson, Richard B., Mitchell, Christina A., and Phillips, Wayne A.
- Subjects
PHOSPHOINOSITIDES ,PROTEIN kinases ,AUTOPHOSPHORYLATION ,PHOSPHORYLATION ,POINT mutation (Biology) ,SERINE - Abstract
Background: The α-isoform of the Type 1A Phosphoinositide 3-kinases (PI3Kα) has protein kinase activity as well as phosphoinositide lipid kinase activity. The best described substrate for its protein kinase activity is its regulatory subunit, p85α, which becomes phosphorylated on Serine 608. Phosphorylation of Serine 608 has been reported to down-regulate its lipid kinase activity. Results: We have assessed whether oncogenic mutants of PI3Kα, which have up-regulated lipid kinase activity, have altered levels of Serine 608 phosphorylation compared to wild type PI3Kα, and whether differential phosphorylation of Serine 608 contributes to increased activity of oncogenic forms of PI3Kα with point mutations in the helical or the kinase domains. Despite markedly increased lipid kinase activity, protein kinase activity was not altered in oncogenic compared to wild type forms of PI3Kα. By manipulating levels of phosphorylation of Serine 608 in vitro, we found no evidence that the protein kinase activity of PI3Kα affects its phosphoinositide lipid kinase activity in either wild-type or oncogenic mutants of PI3Kα. Conclusions: Phosphorylation of p85α S608 is not a significant regulator of wild-type or oncogenic PI3Kα lipid kinase activity. [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
45. Tousled kinase TLK1B mediates chromatinassembly in conjunction with Asf1 regardless ofits kinase activity.
- Author
-
De Benedetti, Arrigo
- Subjects
- *
DNA replication , *DNA synthesis , *CHROMATIN , *CHROMOSOMES , *MOLECULAR chaperones , *PHOSPHORYLATION , *GENES , *MOLECULAR genetics , *GENETICS - Abstract
Background: The Tousled Like Kinases (TLKs) are involved in chromatin dynamics, including DNA replication and repair, transcription, and chromosome segregation. Indeed, the first two TLK1 substrates were identified as the histone H3 and Asf1 (a histone H3/H4 chaperone), which immediately suggested a function in chromatin remodeling. However, despite the straightforward assumption that TLK1 acts simply by phosphorylating its substrates and hence modifying their activity, TLK1 also acts as a chaperone. In fact, a kinase-dead (KD) mutant of TLK1B is functional in stimulating chromatin assembly in vitro. However, subtle effects of Asf1 phosphorylation are more difficult to probe in chromatin assembly assays. Not until very recently was the Asf1 site phosphorylated by TLK1 identified. This has allowed for probing directly the functionality of a site-directed mutant of Asf1 in chromatin assembly assays. Findings: Addition of either wt or non-phosphorylatable mutant Asf1 to nuclear extract stimulates chromatin assembly on a plasmid. Similarly, TLK1B-KD stimulates chromatin assembly and it synergizes in reactions with supplemental Asf1 (wt or non-phosphorylatable mutant). Conclusions: Although the actual function of TLKs as mediators of Asf1 activity cannot be easily studied in vivo, particularly since in mammalian cells there are two TLK genes and two Asf1 genes, we were able to study specifically the stimulation of chromatin assembly in vitro. In such assays, clearly the TLK1 kinase activity was not critical, as neither a non-phosphorylatable Asf1 nor use of the TLK1B-KD impaired the stimulation of nucleosome formation. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
46. Linked decreases in liver kinase B1 and AMP-activated protein kinase activity modulate matrix catabolic responses to biomechanical injury in chondrocytes.
- Author
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Petursson F, Husa M, June R, Lotz M, Terkeltaub R, and Liu-Bryan R
- Subjects
- AMP-Activated Protein Kinase Kinases, Animals, Arthritis, Experimental metabolism, Blotting, Western, Cartilage, Articular metabolism, Cattle, Enzyme-Linked Immunosorbent Assay, Gene Knockdown Techniques, Humans, Immunohistochemistry, Mice, Mice, Inbred C57BL, RNA, Small Interfering, Reverse Transcriptase Polymerase Chain Reaction, Stress, Mechanical, Transfection, AMP-Activated Protein Kinases metabolism, Chondrocytes metabolism, Osteoarthritis metabolism, Protein Serine-Threonine Kinases metabolism
- Abstract
Introduction: AMP-activated protein kinase (AMPK) maintains cultured chondrocyte matrix homeostasis in response to inflammatory cytokines. AMPK activity is decreased in human knee osteoarthritis (OA) chondrocytes. Liver kinase B1 (LKB1) is one of the upstream activators of AMPK. Hence, we examined the relationship between LKB1 and AMPK activity in OA and aging cartilages, and in chondrocytes subjected to inflammatory cytokine treatment and biomechanical compression injury, and performed translational studies of AMPK pharmacologic activation., Methods: We assessed activity (phosphorylation) of LKB1 and AMPKα in mouse knee OA cartilage, in aging mouse cartilage (6 to 24 months), and in chondrocytes after mechanical injury by dynamic compression, via immunohistochemistry or western blot. We knocked down LKB1 by siRNA transfection. Nitric oxide, matrix metalloproteinase (MMP)-3, and MMP-13 release were measured by Griess reaction and ELISA, respectively., Results: Knockdown of LKB1 attenuated chondrocyte AMPK activity, and increased nitric oxide, MMP-3 and MMP-13 release (P <0.05) in response to IL-1β and TNFα. Both LKB1 and AMPK activity were decreased in mouse knee OA and aged knee cartilage, and in bovine chondrocytes after biomechanical injury. Pretreatment of bovine chondrocytes with AMPK activators AICAR and A-769662 inhibited both AMPKα dephosphorylation and catabolic responses after biomechanical injury., Conclusion: LKB1 is required for chondrocyte AMPK activity, thereby inhibiting matrix catabolic responses to inflammatory cytokines. Concurrent loss of LKB1 and AMPK activity in articular chondrocytes is associated with OA, aging and biomechanical injury. Conversely, pharmacologic AMPK activation attenuates catabolic responses to biomechanical injury, suggesting a potentially novel approach to inhibit OA development and progression.
- Published
- 2013
- Full Text
- View/download PDF
47. SU11652 Inhibits tyrosine kinase activity of FLT3 and growth of MV-4-11 cells.
- Author
-
Guo Y, Chen Y, Xu X, Fu X, and Zhao ZJ
- Subjects
- Cell Growth Processes drug effects, Cell Line, Tumor, Humans, Leukemia, Myeloid, Acute enzymology, Leukemia, Myeloid, Acute pathology, Signal Transduction drug effects, fms-Like Tyrosine Kinase 3 metabolism, Indoles pharmacology, Leukemia, Myeloid, Acute drug therapy, Protein Kinase Inhibitors pharmacology, Pyrroles pharmacology, fms-Like Tyrosine Kinase 3 antagonists & inhibitors
- Abstract
Background: FLT3-ITD and FLT3-TKD mutations are frequently found in acute myeloid leukemia (AML). This makes tyrosine kinase FLT3 a highly attractive target for therapeutic drug development. However, effective drugs have not yet emerged. This study is intended to identify and to characterize new FLT3 inhibitors., Methods: By using the protein substrate GST-FLT3S to analyze kinase activity of recombinant proteins carrying the catalytic domain of wild type and mutant forms of FLT3, we screened a chemical library containing 80 known protein kinase inhibitors. We identified SU11652 as a potent FLT3 inhibitor and further employed FLT3-ITD-positive MV- 4-11 cells to study its effects on cell growth, apoptosis, cell cycles, and cell signaling., Results: SU11652 strongly inhibited the activity of wild type, D835Y, and D835H mutant forms of FLT3 with IC50 values of 1.5, 16, and 32 nM, respectively. It effectively blocked the growth of FLT3-ITD -positive MV-4-11 cells at nanomolar concentrations but exhibited much less effects on several other cells which do not carry mutations of FLT3. SU11652 inhibited growth of MV-4-11 cells by inducing apoptosis, causing cell cycle arrest, and blocking activation of the ERK, Akt, and STAT signaling pathways., Conclusion: SU11652 is a potent FLT3 inhibitor which selectively targets FLT3-ITD-positive cells. It should serve as a good candidate for development of therapeutic drugs to treat AML.
- Published
- 2012
- Full Text
- View/download PDF
48. Inhibition of Src kinase activity attenuates amyloid associated microgliosis in a murine model of Alzheimer's disease.
- Author
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Dhawan G and Combs CK
- Subjects
- Alzheimer Disease drug therapy, Amyloid beta-Peptides toxicity, Animals, Animals, Newborn, Cell Line, Cell Line, Transformed, Cells, Cultured, Dasatinib, Enzyme Activation drug effects, Enzyme Activation physiology, Longitudinal Studies, Mice, Mice, Inbred C57BL, Mice, Transgenic, Microglia drug effects, Peptide Fragments toxicity, Protein Kinase Inhibitors pharmacology, Protein Kinase Inhibitors therapeutic use, Pyrimidines pharmacology, Pyrimidines therapeutic use, Thiazoles pharmacology, Thiazoles therapeutic use, Alzheimer Disease enzymology, Amyloid beta-Peptides administration & dosage, Disease Models, Animal, Microglia enzymology, Microglia pathology, Peptide Fragments administration & dosage, src-Family Kinases antagonists & inhibitors, src-Family Kinases physiology
- Abstract
Background: Microglial activation is an important histologic characteristic of the pathology of Alzheimer's disease (AD). One hypothesis is that amyloid beta (Aβ) peptide serves as a specific stimulus for tyrosine kinase-based microglial activation leading to pro-inflammatory changes that contribute to disease. Therefore, inhibiting Aβ stimulation of microglia may prove to be an important therapeutic strategy for AD., Methods: Primary murine microglia cultures and the murine microglia cell line, BV2, were used for stimulation with fibrillar Aβ1-42. The non-receptor tyrosine kinase inhibitor, dasatinib, was used to treat the cells to determine whether Src family kinase activity was required for the Aβ stimulated signaling response and subsequent increase in TNFα secretion using Western blot analysis and enzyme-linked immunosorbent assay (ELISA), respectively. A histologic longitudinal analysis was performed using an AD transgenic mouse model, APP/PS1, to determine an age at which microglial protein tyrosine kinase levels increased in order to administer dasatinib via mini osmotic pump diffusion. Effects of dasatinib administration on microglial and astroglial activation, protein phosphotyrosine levels, active Src kinase levels, Aβ plaque deposition, and spatial working memory were assessed via immunohistochemistry, Western blot, and T maze analysis., Results: Aβ fibrils stimulated primary murine microglia via a tyrosine kinase pathway involving Src kinase that was attenuated by dasatinib. Dasatinib administration to APP/PS1 mice decreased protein phosphotyrosine, active Src, reactive microglia, and TNFα levels in the hippocampus and temporal cortex. The drug had no effect on GFAP levels, Aβ plaque load, or the related tyrosine kinase, Lyn. These anti-inflammatory changes correlated with improved performance on the T maze test in dasatinib infused animals compared to control animals., Conclusions: These data suggest that amyloid dependent microgliosis may be Src kinase dependent in vitro and in vivo. This study defines a role for Src kinase in the microgliosis characteristic of diseased brains and suggests that particular tyrosine kinase inhibition may be a valid anti-inflammatory approach to disease. Dasatinib is an FDA-approved drug for treating chronic myeloid leukemia cancer with a reported ability to cross the blood-brain barrier. Therefore, this suggests a novel use for this drug as well as similar acting molecules.
- Published
- 2012
- Full Text
- View/download PDF
49. Is inhibition of kinase activity the only therapeutic strategy for LRRK2-associated Parkinson's disease?
- Author
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Rudenko IN, Chia R, and Cookson MR
- Subjects
- Animals, Clinical Trials as Topic, Humans, Mutation genetics, Parkinson Disease genetics, Protein Serine-Threonine Kinases chemistry, Protein Serine-Threonine Kinases metabolism, Translational Research, Biomedical, Parkinson Disease drug therapy, Parkinson Disease enzymology, Protein Kinase Inhibitors pharmacology, Protein Kinase Inhibitors therapeutic use, Protein Serine-Threonine Kinases antagonists & inhibitors
- Abstract
Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are a common cause of familial Parkinson's disease (PD). Variation around the LRRK2 locus also contributes to the risk of sporadic PD. The LRRK2 protein contains a central catalytic region, and pathogenic mutations cluster in the Ras of complex protein C terminus of Ras of complex protein (mutations N1437H, R1441G/C and Y1699C) and kinase (G2019S and I2020T) domains. Much attention has been focused on the kinase domain, because kinase-dead versions of mutant LRRK2 are less toxic than kinase-active versions of the same proteins. Furthermore, kinase inhibitors may be able to mimic this effect in mouse models, although the currently tested inhibitors are not completely specific. In this review, we discuss the recent progress in the development of specific LRRK2 kinase inhibitors. We also discuss non-kinase-based therapeutic strategies for LRRK2-associated PD as it is possible that different approaches may be needed for different mutations.
- Published
- 2012
- Full Text
- View/download PDF
50. Continuous requirement of ErbB2 kinase activity for loss of cell polarity and lumen formation in a novel ErbB2/Neu-driven murine cell line model of metastatic breast cancer.
- Author
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Ortega-Cava CF, Raja SM, Laiq Z, Bailey TA, Luan H, Mohapatra B, Williams SH, Ericsson AC, Goswami R, Dimri M, Duan L, Band V, Naramura M, and Band H
- Abstract
Background: Well over a quarter of human breast cancers are ErbB2-driven and constitute a distinct subtype with substantially poorer prognosis. Yet, there are substantial gaps in our understanding of how ErbB2 tyrosine kinase activity unleashes a coordinated program of cellular and extracellular alterations that culminate in aggressive breast cancers. Cellular models that exhibit ErbB2 kinase dependency and can induce metastatic breast cancer in immune competent hosts are likely to help bridge this gap., Materials and Methods: Here, we derived and characterized a cell line model obtained from a transgenic ErbB2/Neu-driven mouse mammary adenocarcinoma., Results: The MPPS1 cell line produces metastatic breast cancers when implanted in the mammary fat pads of immune-compromised as well as syngeneic immune-competent hosts. MPPS1 cells maintain high ErbB2 overexpression when propagated in DFCI-1 or related media, and their growth is ErbB2-dependent, as demonstrated by concentration-dependent inhibition of proliferation with the ErbB kinase inhibitor Lapatinib. When grown in 3-dimensional (3-D) culture on Matrigel, MPPS1 cells predominantly form large irregular cystic and solid structures. Remarkably, low concentrations of Lapatinib led to a switch to regular acinar growth on Matrigel. Immunofluorescence staining of control vs. Lapatinib-treated acini for markers of epithelial polarity revealed that inhibition of ErbB2 signaling led to rapid resumption of normal mammary epithelium-like cell polarity., Conclusions: The strict dependence of the MPPS1 cell system on ErbB2 signals for proliferation and alterations in cell polarity should allow its use to dissect ErbB2 kinase-dependent signaling pathways that promote loss of cell polarity, a key component of the epithelial mesenchymal transition and aggressiveness of ErbB2-driven breast cancers.
- Published
- 2011
- Full Text
- View/download PDF
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