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Xuemei Lin, Gang Wei, Zhijun Huang, Zhongsen Qu, Xinfeng Huang, Hua Xu, Jianjun Liu, Zhixiong Zhuang, Xifei Yang
Mitochondrial dysfunction is involved in neurotoxicity caused by exposure of various chemicals such as copper. However, the effects of long-term low-dose copper exposure on mitochondrial proteome remain unclear. In this study, we found the treatment of copper (0.13ppm copper sulfate in drinking water) for 12 months caused abnormal expression of a total of 13 mitochondrial proteins (7 up-regulated and 6 down-regulated) as revealed by two-dimensional electrophoresis coupled with mass spectrometry in mouse cortex...
October 18, 2016: Toxicology Letters
Hang Chen, Xiuran Niu, Aiai Gao, Sihe Zhang
Objective To investigate the role of mitoch-ondrial signal peptide in guiding enhanced green fluorescent protein-glucose-regulated protein 75 fusion proteins (EGFP-GRP75) into mitochondria. Methods At first, the signal peptide gene and GRP75 domain genes were spliced by overlap extension PCR. Unphosphorylatable mutant T62A/S65A, phospho-mimic mutant T62D/S65D and substrate binding-defective mutant V482F were further created through site-directed mutagenesis PCR. The fusion gene fragments were ligated into pEGFPC1 expression plasmid, respectively...
October 2016: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
Niv Mazkereth, Francesco Rocca, Jennifer-Rose Schubert, Claudia Geisler, Yaron Hillman, Alexander Egner, Zvi Fishelson
Mortalin/GRP75 is a ubiquitously expressed mitochondrial chaperon that is overexpressed in cancer. Mortalin protects cells from complement-dependent cytotoxicity (CDC) and facilitates elimination of the complement C5b-9 complexes from the cell surface. We performed a nanoscopical study aimed at imaging the distribution of the C5b-9 complexes in the plasma membrane and the postulated relocation of mortalin from the mitochondria to the plasma membrane. To gain a resolution of 35nm, the locations of the C5b-9 complex and mortalin were imaged with a STED (Stimulated Emission Depletion) microscope at sub-diffraction resolution...
July 21, 2016: Immunobiology
Yaron Hillman, Niv Mazkereth, Luba Farberov, Noam Shomron, Zvi Fishelson
The impact of microRNAs (miRNAs) known to regulate numerous biologic processes on complement-dependent cytotoxicity (CDC) was investigated in K562 cells. The C5b-9 complex is the executioner of CDC. Cells protect themselves from CDC by C5b-9 elimination, a process involving the mitochondrial chaperone mortalin/GRP75. Potential miR-200 (b and c) and miR-217 regulatory sites were identified in mortalin mRNA. Overexpression of miR-200b/c or miR-217 lowered the expression of mortalin mRNA. miRNA inhibitors for miR-200b, miR-200c, or miR-217 enhanced mortalin mRNA level...
June 15, 2016: Journal of Immunology: Official Journal of the American Association of Immunologists
Hang Chen, Zhihui Gao, Changzheng He, Rong Xiang, Toin H van Kuppevelt, Mattias Belting, Sihe Zhang
Therapeutic macromolecules are internalized into the cell by either clathrin-mediated endocytosis (CME) or clathrin-independent endocytosis (CIE). Although some chaperone proteins play an essential role in CME (e.g. Hsc70 in clathrin uncoating), relatively few of these proteins are functionally involved in CIE. We previously revealed a role for the mitochondrial chaperone protein GRP75 in heparan sulfate proteoglycan (HSPG)-mediated, membrane raft-associated macromolecule endocytosis. However, the mechanism underlying this process remains unclear...
May 1, 2016: Experimental Cell Research
Pavan Kumar Kakumani, Guruprasad R Medigeshi, Inderjeet Kaur, Pawan Malhotra, Sunil K Mukherjee, Raj K Bhatnagar
In recent times, RNAi has emerged as an important defence system that regulates replication of pathogens in host cells. Many RNAi related host factors especially the host miRNAs play important roles in all intrinsic cellular functions, including viral infection. We have been working on identification of mammalian host factors involved in Dengue virus infection. In the present study, we identified Glucose Regulated Protein 75kDa (GRP75), as a host factor that is associated with dicer complex, in particular with HADHA (trifunctional enzyme subunit alpha, mitochondrial), an auxiliary component of dicer complex...
July 15, 2016: Gene
Ainhoa Martínez-Pizarro, Lourdes R Desviat, Magdalena Ugarte, Belén Pérez, Eva Richard
Proper function of endoplasmic reticulum (ER) and mitochondria is crucial for cellular homeostasis, and dysfunction at either site as well as perturbation of mitochondria-associated ER membranes (MAMs) have been linked to neurodegenerative and metabolic diseases. Previously, we have observed an increase in ROS and apoptosis levels in patient-derived fibroblasts with remethylation disorders causing homocystinuria. Here we show increased mRNA and protein levels of Herp, Grp78, IP3R1, pPERK, ATF4, CHOP, asparagine synthase and GADD45 in patient-derived fibroblasts suggesting ER stress and calcium perturbations in homocystinuria...
2016: PloS One
Haidan Jin, Meiying Ji, Liyan Chen, Qixiang Liu, Shuanlong Che, Ming Xu, Zhenhua Lin
BACKGROUND: Mortalin/GRP75 is a ubiquitous mitochondrial chaperone which related to the cytosolic heat shock protein 70 (HSP70), and plays a role in carcinogenesis. This study aims to investigate the Mortalin expression in breast cancer and its correlation with the outcome of the patients with breast cancer. METHODS: A total of 155 invasive ductal carcinoma of breast patients with strict follow-up, 52 ductal carcinoma in situ (DCIS) and 45 adjacent non-tumor breast tissues were selected for immunohistochemical (IHC) staining of Mortalin protein...
2016: Journal of Experimental & Clinical Cancer Research: CR
Song Cao, Yun Liu, Haiying Wang, Xiaowen Mao, Jincong Chen, Jiming Liu, Zhengyuan Xia, Lin Zhang, Xingkui Liu, Tian Yu
Ischemia postconditioning (IPo) is a promising strategy in reducing myocardial ischemia reperfusion (I/R) injury (MIRI), but its specific molecular mechanism is incompletely understood. Langendorff-perfused isolated rat hearts were subjected to global I/R and received IPo in the absence or presence of the mitochondrial ATP-sensitive potassium channel (mitoKATP) blocker 5-hydroxydecanoate (5-HD). Myocardial mitochondria were extracted and mitochondrial comparative proteomics was analyzed. IPo significantly reduces post-ischemic myocardial infarction and improved cardiac function in I/R rat hearts, while 5-HD basically cancelled IPo's myocardial protective effect...
2016: PeerJ
Laurence Booth, Brian Shuch, Thomas Albers, Jane L Roberts, Mehrad Tavallai, Stefan Proniuk, Alexander Zukiwski, Dasheng Wang, Ching-Shih Chen, Don Bottaro, Heath Ecroyd, Iryna O Lebedyeva, Paul Dent
We performed proteomic studies using the GRP78 chaperone-inhibitor drug AR-12 (OSU-03012) as bait. Multiple additional chaperone and chaperone-associated proteins were shown to interact with AR-12, including: GRP75, HSP75, BAG2; HSP27; ULK-1; and thioredoxin. AR-12 down-regulated in situ immuno-fluorescence detection of ATP binding chaperones using antibodies directed against the NH2-termini of the proteins but only weakly reduced detection using antibodies directed against the central and COOH portions of the proteins...
March 15, 2016: Oncotarget
Anne Grünewald, Karolina A Rygiel, Philippa D Hepplewhite, Christopher M Morris, Martin Picard, Doug M Turnbull
OBJECTIVE: To determine the extent of respiratory chain abnormalities and investigate the contribution of mtDNA to the loss of respiratory chain complexes (CI-IV) in the substantia nigra (SN) of idiopathic Parkinson disease (IPD) patients at the single-neuron level. METHODS: Multiple-label immunofluorescence was applied to postmortem sections of 10 IPD patients and 10 controls to quantify the abundance of CI-IV subunits (NDUFB8 or NDUFA13, SDHA, UQCRC2, and COXI) and mitochondrial transcription factors (TFAM and TFB2M) relative to mitochondrial mass (porin and GRP75) in dopaminergic neurons...
March 2016: Annals of Neurology
Eleni Stamoula, Theofanis Vavilis, Eleni Aggelidou, Aikaterini Kaidoglou, Angeliki Cheva, Kyriakos Mellidis, Antigone Lazou, Costas Haitoglou, Maria Albani, Aristeidis Kritis
BACKGROUND/AIMS: Increasing amounts of the neurotransmitter glutamate are associated with excitotoxicity, a phenomenon related both to homeostatic processes and neurodegenerative diseases such as multiple sclerosis. METHODS: PC12 cells (rat pheochromocytoma) were treated with various concentrations of the non-essential amino acid glutamate for 0.5-24 hours. The effect of glutamate on cell morphology was monitored with electron microscopy and haematoxylin-eosin staining...
2015: Cellular Physiology and Biochemistry
Mohammed Gagaoua, E M Claudia Terlouw, Abdelghani Boudjellal, Brigitte Picard
The development of proteomic biomarkers for meat tenderness remains an important challenge. The present study used Longissimus thoracis (LT) and Semitendinosus (ST) muscles of young bulls of three continental breeds (Aberdeen Angus, Blond d'Aquitaine and Limousin) to i) identify cellular pathways robustly related with meat tenderness, using potential protein biomarkers and ii) describe biochemical mechanisms underlying muscle to meat conversion. Correlation networks reveal robust correlations, i.e. present for at least two breeds, between potential meat tenderness biomarkers...
October 14, 2015: Journal of Proteomics
Duangnate Isarangkul, Suthep Wiyakrutta, Kanchana Kengkoom, Onrapak Reamtong, Sumate Ampawong
The pathogenesis of congenital hydronephrosis in laboratory animals has been studied for many years, yet little is known about the underlying mechanism of this disease. In this study, we investigated a MS-based comparative proteomics approach to characterize the differently expressed proteins between kidney tissue samples of ICR/Mlac-hydro and wild-type mice. Interestingly, proteomic results exhibited several mitochondrial protein alterations especially the up-regulation of 60 kDa heat shock protein (Hsp60), stress-70 protein (GRP75) dysfunction, and down-regulation of voltage-dependent anion-selective channel protein 1 (VDAC-1)...
2015: International Journal of Clinical and Experimental Medicine
Theofanis Vavilis, Nikoleta Delivanoglou, Eleni Aggelidou, Eleni Stamoula, Kyriakos Mellidis, Aikaterini Kaidoglou, Angeliki Cheva, Chryssa Pourzitaki, Katerina Chatzimeletiou, Antigone Lazou, Maria Albani, Aristeidis Kritis
Hypoxia is the lack of sufficient oxygenation of tissue, imposing severe stress upon cells. It is a major feature of many pathological conditions such as stroke, traumatic brain injury, cerebral hemorrhage, perinatal asphyxia and can lead to cell death due to energy depletion and increased free radical generation. The present study investigates the effect of hypoxia on the unfolded protein response of the cell (UPR), utilizing a 16-h oxygen-glucose deprivation protocol (OGD) in a PC12 cell line model. Expression of glucose-regulated protein 78 (GRP78) and glucose-regulated protein 94 (GRP94), key players of the UPR, was studied along with the expression of glucose-regulated protein 75 (GRP75), heat shock cognate 70 (HSC70), and glyceraldehyde 3-phosphate dehydrogenase, all with respect to the cell death mechanism(s)...
July 2016: Cellular and Molecular Neurobiology
Gry Aune Westergaard Hansen, Maja Ludvigsen, Christian Jacobsen, Claudia Cangemi, Lars Melholt Rasmussen, Henrik Vorum, Bent Honoré
Affinity purification, immunoprecipitation, gel electrophoresis and mass spectrometry were used to identify fibulin-1C, C1 esterase inhibitor and glucose regulated protein 75, grp75, as binding partners of the CREC proteins, calumenin and reticulocalbin. Surface plasmon resonance was used to verify the interaction of all three proteins with each of the CREC proteins. Fibulin-1C interacts with calumenin and reticulocalbin with an estimated dissociation constant around 50-60 nM. The interaction, at least for reticulocalbin, was not dependent upon the presence of Ca2+...
2015: PloS One
Feng-Tao Liu, Yan Chen, Yu-Jie Yang, Ling Yang, Mei Yu, Jian Zhao, Jian-Jun Wu, Fang Huang, Wen Liu, Zheng-Tong Ding, Jian Wang
Mutations and excessive accumulation of α-synuclein (α-syn) can lead to the degeneration of dopaminergic neurons, indicating a pivotal role of α-syn in the pathogenesis of Parkinson's disease (PD). Although how α-syn contributes to PD is still elusive, mitochondrial impairments have been reported to be implicated in. Mortalin, a molecular chaperone mainly located in mitochondria, has been linked to the pathogenesis of PD in recent studies. Moreover, some proteomics studies indicate that mortalin is associated with PD-related proteins, including α-syn...
April 16, 2015: Brain Research
Renu Wadhwa, Jihoon Ryu, Hyo Min Ahn, Nishant Saxena, Anupama Chaudhary, Chae-Ok Yun, Sunil C Kaul
Mortalin/mtHsp70/Grp75 (mot-2), a heat shock protein 70 family member, is an essential chaperone, enriched in cancers, and has been shown to possess pro-proliferative and anti-apoptosis functions. An allelic form of mouse mortalin (mot-1) that differs by two amino acids, M618V and G624R, in the C terminus substrate-binding domain has been reported. Furthermore, genome sequencing of mortalin from Parkinson disease patients identified two missense mutants, R126W and P509S. In the present study, we investigated the significance of these mutations in survival, proliferation, and oxidative stress tolerance in human cells...
March 27, 2015: Journal of Biological Chemistry
Gaoming Zhang, Mei Han, Xiaodong Wang, Aiping Xiao
Glucose-regulated protein 75 (GRP75), a member of the heat-shock protein 70 family, is known to protect cells from stress-induced injury. However, information regarding its distribution and possible function in the retina is limited. In this study, we performed an optic nerve crush (ONC) model in adult rats and found that GRP75 was significantly upregulated in the retina after ONC. Double immunofluorescent staining revealed that GRP75 was localized in the retinal ganglion cells (RGCs). We also examined the expression profile of active caspase3, whose change was correlated with the expression profile of GRP75...
June 2015: Journal of Molecular Neuroscience: MN
Niku K J Oksala, F Güler Ekmekçi, Ergi Ozsoy, Serife Kirankaya, Tarja Kokkola, Güzin Emecen, Jani Lappalainen, Kai Kaarniranta, Mustafa Atalay
Heat shock proteins (HSPs), originally identified as heat-inducible gene products, are a family of highly conserved proteins that respond to a wide variety of stress including oxidative stress. Although both acute and chronic oxidative stress have been well demonstrated to induce HSP responses, little evidence is available whether increased HSP levels provide enhanced protection against oxidative stress under elevated yet sublethal temperatures. We studied relationships between oxidative stress and HSPs in a physiological model by using Garra rufa (doctor fish), a fish species naturally acclimatized to different thermal conditions...
2014: Redox Biology
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