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https://www.readbyqxmd.com/read/29053695/targeting-cysteine-thiols-for-in-vitro-site-specific-glycosylation-of-recombinant-proteins
#1
Yoo Jung Choi, Jinhui Zhu, Steve Chung, Naveed Siddiqui, Qingping Feng, Peter B Stathopulos
Stromal interaction molecule-1 (STIM1) is a type-I transmembrane protein located on the endoplasmic reticulum (ER) and plasma membranes (PM). ER-resident STIM1 regulates the activity of PM Orai1 channels in a process known as store operated calcium (Ca(2+)) entry which is the principal Ca(2+) signaling process that drives the immune response. STIM1 undergoes post-translational N-glycosylation at two luminal Asn sites within the Ca(2+) sensing domain of the molecule. However, the biochemical, biophysical, and structure biological effects of N-glycosylated STIM1 were poorly understood until recently due to an inability to readily obtain high levels of homogeneous N-glycosylated protein...
October 4, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29051492/calmodulin-dissociates-the-stim1-orai1-complex-and-stim1-oligomers
#2
Xin Li, Guangyan Wu, Yin Yang, Shijuan Fu, Xiaofen Liu, Huimin Kang, Xue Yang, Xun-Cheng Su, Yuequan Shen
Store-operated calcium entry (SOCE) is a major pathway for calcium ions influx into cells and has a critical role in various cell functions. Here we demonstrate that calcium-bound calmodulin (Ca(2+)-CaM) binds to the core region of activated STIM1. This interaction facilitates slow Ca(2+)-dependent inactivation after Orai1 channel activation by wild-type STIM1 or a constitutively active STIM1 mutant. We define the CaM-binding site in STIM1, which is adjacent to the STIM1-Orai1 coupling region. The binding of Ca(2+)-CaM to activated STIM1 disrupts the STIM1-Orai1 complex and also disassembles STIM1 oligomer...
October 19, 2017: Nature Communications
https://www.readbyqxmd.com/read/29048678/suppression-of-stim1-inhibits-the-migration-and-invasion-of-human-prostate-cancer-cells-and-is-associated-with-pi3k-akt-signaling-inactivation
#3
Yibin Zhou, Peng Gu, Jian Li, Feng Li, Jin Zhu, Peng Gao, Yachen Zang, Yongchang Wang, Yuxi Shan, Dongrong Yang
Store-operated calcium entry (SOCE) plays an important role in the invasion and migration of cancer cells. Stromal-interacting molecule 1 (STIM1) is a critical component in the SOCE. STIM1 has been attracting more and more attention due to its oncogenic potential. STIM1 inhibition suppresses cell proliferation, migration and invasion in a variety of cancer models both in vitro and in vivo. However, the role of STIM1 in prostate carcinogenesis, in particular, in tumor migration and invasion is unclear. Herein, we downregulated STIM1 in prostate cancer cells by lentivirus-mediated short hairpin (shRNA), and then studied its impacts on cell migration and invasion...
September 18, 2017: Oncology Reports
https://www.readbyqxmd.com/read/28994003/muscarinic-acetylcholine-receptors-potentiate-5-adenosine-monophosphate-activated-protein-kinase-stimulation-and-glucose-uptake-triggered-by-thapsigargin-induced-store-operated-ca-2-entry-in-human-neuroblastoma-cells
#4
Maria C Olianas, Simona Dedoni, Pierluigi Onali
The 5'-adenosine monophosphate-activated protein kinase (AMPK) is a key regulator of the cellular energy metabolism and may induce either cell survival or death. We previously reported that in SH-SY5Y human neuroblastoma cells stimulation of muscarinic acetylcholine receptors (mAChRs) activate AMPK by triggering store-operated Ca(2+) entry (SOCE). However, whether mAChRs may control AMPK activity by regulating additional mechanisms beyond SOCE remains to be investigated. In the present study we examined the effects of mAChRs on AMPK when SOCE was induced by the sarco-endoplasmic reticulum Ca(2+)-ATPase inhibitor thapsigargin...
October 9, 2017: Neurochemical Research
https://www.readbyqxmd.com/read/28972171/inhibition-of-the-sarco-endoplasmic-reticulum-er-ca2-atpase-by-thapsigargin-analogs-induces-cell-death-via-er-ca2-depletion-and-the-unfolded-protein-response
#5
Pankaj Sehgal, Paula Szalai, Claus Olesen, Helle A Praetorius, Poul Nissen, Søren Brøgger Christensen, Nikolai Engedal, Jesper Vuust Møller
Calcium (Ca2+) is a fundamental regulator of cell signaling and function. Thapsigargin (Tg) blocks the sarco/endoplasmic reticulum (ER) Ca2+-ATPase (SERCA), disrupts Ca2+ homeostasis, and causes cell death. However, the exact mechanisms whereby SERCA-inhibition induces cell death are incompletely understood. Here, we report that low (0.1 μM) concentrations of Tg and Tg analogs with various long-chain substitutions at the O(8) position extensively inhibit SERCA1a-mediated Ca2+ transport. We also found that in both prostate and breast cancer cells, exposure to Tg or Tg analogs for 1 day caused extensive drainage of the ER Ca2+ stores...
September 29, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28966111/cationic-liposomes-suppress-intracellular-calcium-ion-concentration-increase-via-inhibition-of-pi3-kinase-pathway-in-mast-cells
#6
Yoshikazu Inoh, Aki Haneda, Satoshi Tadokoro, Satoru Yokawa, Tadahide Furuno
Cationic liposomes are commonly used as vectors to effectively introduce foreign genes (antisense DNA, plasmid DNA, siRNA, etc.) into target cells. Cationic liposomes are also known to affect cellular immunocompetences such as the mast cell function in allergic reactions. In particular, we previously showed that the cationic liposomes bound to the mast cell surface suppress the degranulation induced by cross-linking of high affinity IgE receptors in a time- and dose-dependent manner. This suppression is mediated by impairment of the sustained level of intracellular Ca(2+) concentration ([Ca(2+)]i) via inhibition of store-operated Ca(2+) entry (SOCE)...
September 29, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28955036/optimizing-the-fragment-complementation-of-apex2-for-detection-of-specific-protein-protein-interactions-in-live-cells
#7
Miaomiao Xue, Junjie Hou, Linlin Wang, Dongwan Cheng, Jingze Lu, Li Zheng, Tao Xu
Dynamic protein-protein interactions (PPIs) play crucial roles in cell physiological processes. The protein-fragment complementation (PFC) assay has been developed as a powerful approach for the detection of PPIs, but its potential for identifying protein interacting regions is not optimized. Recently, an ascorbate peroxidase (APEX2)-based proximity-tagging method combined with mass spectrometry was developed to identify potential protein interactions in live cells. In this study, we tested whether APEX2 could be employed for PFC...
September 27, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28935720/suppression-of-store-operated-ca-2-entry-by-activation-of-gper-contribution-to-a-clamping-effect-on-endothelial-ca-2-signaling
#8
Lara E Terry, Mark VerMeer, Jennifer Giles, Quang-Kim Tran
The G Protein-coupled Estrogen Receptor 1 (GPER, formerly a.k.a. GPR30) modulates many Ca(2+)-dependent activities in endothelial cells. However, the underlying mechanisms are poorly understood. We recently reported that GPER acts to prolong cytoplasmic Ca(2+) signals by interacting with and promoting inhibitory phosphorylation of the plasma membrane Ca(2+)-ATPase. In this study, we examined the role of GPER activation in modulating store-operated Ca(2+) entry (SOCE) via effects on the Stromal Interaction Molecule 1 (STIM1)...
September 21, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28930597/the-store-operated-calcium-channels-in-cancer-metastasis-from-cell-migration-invasion-to-metastatic-colonization
#9
Pingli Mo, Shengyu Yang
Store-operated calcium entry (SOCE) is the predominant calcium entry mechanism in most cancer cells. SOCE is mediated by the endoplasmic reticulum calcium sensor STIMs (STIM1 and 2) and plasma membrane channel forming unit Orais (Orai 1-3). In recent years there is increasing evidence indicating that SOCE in cancer cells is dysregulated to promote cancer cell migration, invasion and metastasis. The overexpression of STIM and Orai proteins has been reported to correlate with the metastatic progression of various cancers...
January 1, 2018: Frontiers in Bioscience (Landmark Edition)
https://www.readbyqxmd.com/read/28921392/resveratrol-attenuates-high-glucose-induced-endothelial-cell-apoptosis-via-mediation-of-store-operated-calcium-entry
#10
Ting Lu, Dayan Zhou, Pan Gao, Liangyi Si, Qiang Xu
The aim of this study was to evaluate the influence of resveratrol on HG-induced calcium entry in islet microvascular (MS-1) endothelial cells. MS-1 cells were pretreated with resveratrol or 2-APB (an inhibitor of store-operated calcium entry) and then incubated with high glucose. Cell viability was determined using the cell counting kit-8 method. Reactive oxygen species, endothelial apoptosis, and NO production were detected by DHE probe, TUNEL detection, and nitrate reductase assay kit. Protein levels of SOCE were detected by western blotting...
September 18, 2017: Molecular and Cellular Biochemistry
https://www.readbyqxmd.com/read/28919480/cholesterol-modulates-the-cellular-localization-of-orai1-channels-and-its-disposition-among-membrane-domains
#11
A Bohórquez-Hernández, Enrico Gratton, Jonathan Pacheco, Alexander Asanov, Luis Vaca
Store Operated Calcium Entry (SOCE) is one of the most important mechanisms for calcium mobilization in to the cell. Two main proteins sustain SOCE: STIM1 that acts as the calcium sensor in the endoplasmic reticulum (ER) and Orai1 responsible for calcium influx upon depletion of ER. There are many studies indicating that SOCE is modulated by the cholesterol content of the plasma membrane (PM). However, a myriad of questions remain unanswered concerning the precise molecular mechanism by which cholesterol modulates SOCE...
December 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28912430/stim1-dependent-ca-2-signaling-regulates-podosome-formation-to-facilitate-cancer-cell-invasion
#12
Yun-Wen Chen, Yih-Fung Chen, Wen-Tai Chiu, Hong-Chen Chen, Meng-Ru Shen
The clinical significance of STIM proteins and Orai Ca(2+) channels in tumor progression has been demonstrated in different types of cancers. Podosomes are dynamic actin-rich cellular protrusions that facilitate cancer cell invasiveness by degrading extracellular matrix. Whether STIM1-dependent Ca(2+) signaling facilitates cancer cell invasion through affecting podosome formation remains unclear. Here we show that the invasive fronts of cancer tissues overexpress STIM1, accompanied by active store-operated Ca(2+) entry (SOCE)...
September 14, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28900935/metabolic-disorders-and-cancer-hepatocyte-store-operated-ca-2-channels-in-nonalcoholic-fatty-liver-disease
#13
Eunüs S Ali, Grigori Y Rychkov, Greg J Barritt
In steatotic hepatocytes, intracellular Ca(2+) homeostasis is substantially altered compared to normal. Decreased Ca(2+) in the endoplasmic reticulum (ER) can lead to ER stress, an important mediator of the progression of liver steatosis to nonalcoholic steatohepatitis, type 2 diabetes, and hepatocellular carcinoma. Store-operated Ca(2+) channels (SOCs) in hepatocytes are composed principally of Orai1 and STIM1 proteins. Their main role is the maintenance of adequate Ca(2+) in the lumen of the ER. In steatotic hepatocytes, store-operated Ca(2+) entry (SOCE) is substantially inhibited...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/28900934/fertility-store-operated-ca-2-entry-in-germ-cells-role-in-egg-activation
#14
Zoltan Machaty, Chunmin Wang, Kiho Lee, Lu Zhang
At the time of fertilization, the sperm activates the egg and induces embryonic development by triggering an elevation in the egg's intracellular free Ca(2+) concentration. In mammals the initial Ca(2+) rise is followed by a series of repetitive Ca(2+) transients (known as oscillations) that last for several hours. Although the source of Ca(2+) during the signaling process is primarily the egg's smooth endoplasmic reticulum, the oscillations stop in the absence of extracellular Ca(2+) indicating that a Ca(2+) influx across the plasma membrane is essential to sustain them...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/28900931/cardiac-remodeling-and-disease-current-understanding-of-stim1-orai1-mediated-store-operated-ca-2-entry-in-cardiac-function-and-pathology
#15
Fiona Bartoli, Jessica Sabourin
For a long time, Ca(2+) entry into cardiomyocytes was considered the sole domain of the L-type Ca(2+) channel. Recently, STIM1/Orai1-mediated store-operated Ca(2+) entry has been also reported to participate to Ca(2+) influx in cardiac cells and has emerged as a key player to alter Ca(2+) in the cardiomyocyte. In this review, we will highlight accumulated knowledge about the presence and the potential contribution of STIM1/Orai1-dependent SOCE to cardiac function and its role in the cardiac pathogenesis. Overall, even if STIM1/Orai1 proteins are present in the heart, contradictory results have been reported regarding their contribution to cardiac physiology and pathology, pointing out the necessity of further investigations, a major challenge over the coming years...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/28900930/cardiac-remodeling-and-disease-soce-and-trpc-signaling-in-cardiac-pathology
#16
Petra Eder
TRPC channels have been suggested as potential candidates mediating store-operated Ca(2+) entry (SOCE) in cardiomyocytes. There is increasing evidence that the TRPC isoforms TRPC1 and TRPC4 might fulfill the function as SOCs, in concert with or in parallel to the key players of SOCE, Orai1, and STIM1. Several other isoforms, e.g., TRPC3, TRPC6, and TRPC7, might rather associate to receptor-activated diacylglycerol (DAG)-sensitive ion channels. However, the exact activation mode has not been elucidated yet, given the characteristic of TRPC channels to heteromerize to unpredictable ion channel assemblies...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/28900928/cardiovascular-and-hemostatic-disorders-soce-and-ca-2-handling-in-platelet-dysfunction
#17
Jose J Lopez, Gines M Salido, Juan A Rosado
Among the Ca(2+) entry mechanisms in platelets, store-operated Ca(2+) entry (SOCE) plays a prominent role as it is necessary to achieve full activation of platelet functions and replenish intracellular Ca(2+) stores. In platelets, as in other non-excitable cells, SOCE has been reported to involve the activation of plasma membrane channels by the ER Ca(2+) sensor STIM1. Despite electrophysiological studies are not possible in human platelets, indirect analyses have revealed that the Ca(2+)-permeable channels involve Orai1 and, most likely, TRPC1 subunits...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/28900927/cardiovascular-and-hemostatic-disorders-role-of-stim-and-orai-proteins-in-vascular-disorders
#18
Jyoti Tanwar, Mohamed Trebak, Rajender K Motiani
Store-operated Ca(2+) entry (SOCE) mediated by STIM and Orai proteins is a highly regulated and ubiquitous signaling pathway that plays an important role in various cellular and physiological functions. Endoplasmic reticulum (ER) serves as the major site for intracellular Ca(2+) storage. Stromal Interaction Molecule 1/2 (STIM1/2) sense decrease in ER Ca(2+) levels and transmits the message to plasma membrane Ca(2+) channels constituted by Orai family members (Orai1/2/3) resulting in Ca(2+) influx into the cells...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/28900926/immunological-disorders-regulation-of-ca-2-signaling-in-t-lymphocytes
#19
Sonal Srikanth, Jin Seok Woo, Zuoming Sun, Yousang Gwack
Engagement of T cell receptors (TCRs) with cognate antigens triggers cascades of signaling pathways in helper T cells. TCR signaling is essential for the effector function of helper T cells including proliferation, differentiation, and cytokine production. It also modulates effector T cell fate by inducing cell death, anergy (nonresponsiveness), exhaustion, and generation of regulatory T cells. One of the main axes of TCR signaling is the Ca(2+)-calcineurin-nuclear factor of activated T cells (NFAT) signaling pathway...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/28900920/the-role-of-mitochondria-in-the-activation-maintenance-of-soce-membrane-contact-sites-as-signaling-hubs-sustaining-store-operated-ca-2-entry
#20
Nicolas Demaurex, Daniele Guido
Store-operated Ca(2+) entry (SOCE) is a cell signaling pathway essential for immune and muscle function controlled by dynamic interactions between Ca(2+)-sensing STIM proteins on the endoplasmic reticulum (ER) and Ca(2+)-permeable ORAI channels on the plasma membrane (PM). STIM-ORAI interactions occur at membrane contact sites (MCS), evolutionarily conserved cellular structures characterized by the close apposition (10-20 nm) between the ER and target membranes that facilitate the exchange of lipids by non-vesicular transport mechanisms...
2017: Advances in Experimental Medicine and Biology
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