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Recombinant hsp70

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https://www.readbyqxmd.com/read/29140786/selection-of-an-anticalin%C3%A2-against-the-membrane-form-of-hsp70-via-bacterial-surface-display-and-its-theranostic-application-in-tumour-models
#1
Lars Friedrich, Petra Kornberger, Claudia T Mendler, Gabriele Multhoff, Markus Schwaiger, Arne Skerra
We describe the selection of Anticalins against a common tumour surface antigen, human Hsp70, using functional display on live E. coli cells as fusion with a truncated EspP autotransporter. While found intracellularly in normal cells, Hsp70 is frequently exposed in a membrane-bound state on the surface of tumour cells and, even more pronounced, in metastases or after radiochemotherapy. Employing a recombinant Hsp70 fragment comprising residues 383-548 as target, Anticalins were selected from a naive bacterial library...
November 27, 2017: Biological Chemistry
https://www.readbyqxmd.com/read/29127260/an-improved-brome-mosaic-virus-silencing-vector-greater-insert-stability-and-more-extensive-vigs
#2
Xin Shun Ding, Stephen W Mannas, Bethany A Bishop, Xiaolan Rao, Mitchell Lecoultre, Soonil Kwon, Richard S Nelson
Virus-induced gene silencing (VIGS) is used extensively for gene function studies in plants. VIGS is inexpensive and rapid compared with silencing conducted through stable transformation, but many virus silencing vectors, especially in grasses, induce only transient silencing phenotypes. A major reason for transient phenotypes is instability of the foreign gene fragment (insert) in the vector during VIGS. Here we report the development of a Brome mosaic virus (BMV)-based vector that better maintains inserts through modification of the original BMV vector RNA sequence...
November 10, 2017: Plant Physiology
https://www.readbyqxmd.com/read/29107750/molecular-cloning-prokaryotic-expression-purification-structural-studies-and-functional-implications-of-heat-shock-protein-70-hsp70-from-rutilus-frisii-kutum
#3
Zohreh Jahangirizadeh, Hossein Ghafouri, Reza H Sajedi, Sajjad Sarikhan, Majid Taghdir, Reyhaneh Sariri
A novel Hsp70 chaperone from Rutilus frisii kutum was identified, cloned, expressed, purified and its functional characteristics revealed. The 3D structure of Hsp70 from Rutilus kutum was constructed using the crystal structure of E. coli Hsp70 as the template, with 47% sequence identity. The in vitro ATPase activity assay after 60minutes, ATP hydrolysis of purified recombinant Hsp70 (8μM) was improved by binding to denatured thermally luciferase (3μM) about 2.5-fold compared with that of Hsp70 alone. Based on the results, it was found that the purified Hsp70 chaperone was able to considerably suppress heat-induced aggregation of luciferase by binding to DnaJ co-chaperone (5μM) more than 70% after 10min at 42°C...
October 28, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/29102627/the-anti-stresses-capability-of-grp78-in-penaeus-monodon-evidence-from-in-vitro-and-in-vivo-studies
#4
Lili Li, Pengfei Wang, Chao Zhao, Lihua Qiu
Glucose-regulated protein 78 (GRP78) is an important member of the heat shock protein 70 (HSP70) family, which plays important roles in the response to stimulation from various external environmental stimuli. In this study, the full-length cDNA of GRP78 (PmGRP78) was cloned from Penaeus monodon. The qRT-PCR analysis showed that PmGRP78 was ubiquitously expressed in all the tested shrimp tissues, and the expression pattern of PmGRP78 in the gill and hepatopancreas tissues after a pH challenge, osmotic stress, and heavy metal exposure was also detected...
November 2, 2017: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/29067554/extracellular-hsp70-induces-inflammation-and-modulates-lps-lta-stimulated-inflammatory-response-in-thp-1-cells
#5
Andrea Hulina, Marija Grdić Rajković, Daniela Jakšić Despot, Dubravko Jelić, Ana Dojder, Ivana Čepelak, Lada Rumora
Extracellular Hsp70 (eHsp70) can act as damage-associated molecular pattern (DAMP) via Toll-like receptors TLR2 and TLR4, and stimulate immune and inflammatory responses leading to sterile inflammation and propagation of already existing inflammation. It was found elevated in the blood of patients with chronic obstructive pulmonary disease (COPD), who might suffer occasional bacterial colonizations and infections. We used a monocytic THP-1 cell line as a cellular model of systemic compartment of COPD to assess inflammatory effects of eHsp70 when present alone or together with bacterial products lypopolysaccharide (LPS) and lypoteichoic acid (LTA)...
October 24, 2017: Cell Stress & Chaperones
https://www.readbyqxmd.com/read/28951871/recombinant-vaccinia-viruses-coding-transgenes-of-apoptosis-inducing-proteins-enhance-apoptosis-but-not-immunogenicity-of-infected-tumor-cells
#6
Olga Koval, Galina Kochneva, Anastasiya Tkachenko, Olga Troitskaya, Galina Sivolobova, Antonina Grazhdantseva, Anna Nushtaeva, Elena Kuligina, Vladimir Richter
Genetic modifications of the oncolytic vaccinia virus (VV) improve selective tumor cell infection and death, as well as activation of antitumor immunity. We have engineered a double recombinant VV, coding human GM-CSF, and apoptosis-inducing protein apoptin (VV-GMCSF-Apo) for comparing with the earlier constructed double recombinant VV-GMCSF-Lact, coding another apoptosis-inducing protein, lactaptin, which activated different cell death pathways than apoptin. We showed that both these recombinant VVs more considerably activated a set of critical apoptosis markers in infected cells than the recombinant VV coding GM-CSF alone (VV-GMCSF-dGF): these were phosphatidylserine externalization, caspase-3 and caspase-7 activation, DNA fragmentation, and upregulation of proapoptotic protein BAX...
2017: BioMed Research International
https://www.readbyqxmd.com/read/28948857/neuroprotective-effect-of-secreted-factors-from-human-adipose-stem-cells-in-a-rat-stroke-model
#7
Han Gil Seo, Youbin Yi, Byung-Mo Oh, Nam-Jong Paik
Objectives Recent evidence shows that stem cells exert neuroprotective effect through the secretion of immune modulatory, neurotrophic factors. We aimed to assess the neuroprotective effect of selected recombinant factors (RFs) detected in human adipose stem cell (hASC)-conditioned medium (CM), in a rat ischemic stroke model. Methods Ischemic stroke was induced in Sprague-Dawley rats using 2 h transient middle cerebral artery occlusion (MCAO). One hour after reperfusion, the vehicle (Dulbecco's modified Eagle medium; DMEM), concentrated CM, and selected RFs mixed with DMEM were administered intracerebroventricularly to each group (N = 14, 15, and 16, respectively)...
September 26, 2017: Neurological Research
https://www.readbyqxmd.com/read/28928431/tumor-induces-muscle-wasting-in-mice-through-releasing-extracellular-hsp70-and-hsp90
#8
Guohua Zhang, Zhelong Liu, Hui Ding, Yong Zhou, Hoang Anh Doan, Ka Wai Thomas Sin, Zhiren J Zhu, Rene Flores, Yefei Wen, Xing Gong, Qingyun Liu, Yi-Ping Li
Cachexia, characterized by muscle wasting, is a major contributor to cancer-related mortality. However, the key cachexins that mediate cancer-induced muscle wasting remain elusive. Here, we show that tumor-released extracellular Hsp70 and Hsp90 are responsible for tumor's capacity to induce muscle wasting. We detected high-level constitutive release of Hsp70 and Hsp90 associated with extracellular vesicles (EVs) from diverse cachexia-inducing tumor cells, resulting in elevated serum levels in mice. Neutralizing extracellular Hsp70/90 or silencing Hsp70/90 expression in tumor cells abrogates tumor-induced muscle catabolism and wasting in cultured myotubes and in mice...
September 19, 2017: Nature Communications
https://www.readbyqxmd.com/read/28919581/antitumor-response-to-a-codon-optimized-hpv-16-e7-hsp70-fusion-antigen-dna-vaccine
#9
Hoorieh Soleimanjahi, Hadi Razavinikoo, Fatemeh Fotouhi, Abdollah Ardebili
BACKGROUND: Vaccines based on virus-like particles are effective against Human Papilloma Virus (HPV) infection; however, they have not shown a therapeutic effect against HPV-associated diseases. New immunotherapy strategies based on immune responses against tumor antigens can positively affect the clearance of HPV-associated lesions. OBJECTIVE: To generate two therapeutic fusion DNA vaccines (optimizedE7/mouseHSP70 and wildE7/mouseHSP70) to induce antitumor specific responses in mice models...
September 2017: Iranian Journal of Immunology: IJI
https://www.readbyqxmd.com/read/28893913/il17a-deficient-mice-are-highly-susceptible-to-toxoplasma-gondii-infection-due-to-excessively-induced-t-gondii-hsp70-and-ifn-%C3%AE-production
#10
Masataka Moroda, Masaya Takamoto, Yoichiro Iwakura, Jun Nakayama, Fumie Aosai
IL-17A is known to be involved in the host defense against pathogens and pathogenesis of autoimmune diseases. Previously, we showed that excessive IFN-γ plays an important role in the pathogenesis of lethal effect of Toxoplasma gondii (T. gondii) by inducing anaphylactic responses. In this report, we examine the effects of an IL-17A deficiency on murine host defense against oral T. gondii infection. IL-17A-deficient C57BL/6 (B6) mice exhibited higher mortality than wild type (WT) mice to T. gondii at the acute phase of infection...
September 11, 2017: Infection and Immunity
https://www.readbyqxmd.com/read/28819167/artificial-dnaj-protein-for-protein-production-and-conformational-diseases
#11
Akinori Hishiya, Keizo Koya
For secreted proteins, proper protein folding is essential not only for biological function but also for secretion itself. Proteins with folding problems are trapped in the endoplasmic reticulum (ER) and are eventually degraded in the cytoplasm. In this study, we exploited co-expression of an artificial fusion protein, based on the sequence of a DnaJ protein, which could interact as co-chaperones in the Hsp70-based protein-folding system, with target recombinant secreted proteins to enhance their production and secretion...
August 17, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28759972/molecular-mechanisms-underlying-neuroprotective-effect-of-intranasal-administration-of-human-hsp70-in-mouse-model-of-alzheimer-s-disease
#12
Michail B Evgen'ev, George S Krasnov, Inna V Nesterova, David G Garbuz, Vadim L Karpov, Alexey V Morozov, Anastasiya V Snezhkina, Alexander N Samokhin, Alexander Sergeev, Alexei M Kulikov, Natalia V Bobkova
Heat shock protein 70, encoded by the HSPA1A gene in humans, is a key component of the machinery that protects neuronal cells from various stress conditions and whose production significantly declines during the course of aging and as a result of several neurodegenerative diseases. Herein, we investigated whether sub-chronic intranasal administration of exogenous Hsp70 (eHsp70) exerts a neuroprotective effect on the temporal cortex and areas of the hippocampus in transgenic 5XFAD mice, a model of Alzheimer's disease...
2017: Journal of Alzheimer's Disease: JAD
https://www.readbyqxmd.com/read/28745541/chaperone-proteins-as-single-component-reagents-to-assess-antibody-nonspecificity
#13
Ryan L Kelly, James C Geoghegan, Jared Feldman, Tushar Jain, Monique Kauke, Doris Le, Jessie Zhao, K Dane Wittrup
Early stage assays that evaluate monoclonal antibody drug-like properties serve as valuable tools for selection of lead candidates. One liability for clinical development, off-target reactivity, is often assessed by binding to a mixture or panel of noncognate proteins. While robust, these mixes are often ill-defined, and can suffer from issues such as lot-to-lot variability. In this study, we discovered in immunoprecipitation experiments that certain chaperones are present in one of these mixtures;we then explored the use of recombinant chaperone proteins as well-characterized agents to predict antibody nonspecificity...
October 2017: MAbs
https://www.readbyqxmd.com/read/28679534/hsp70-s-rna-binding-and-mrna-stabilizing-activities-are-independent-of-its-protein-chaperone-functions
#14
Aparna Kishor, Elizabeth J F White, Aerielle E Matsangos, Zisui Yan, Bishal Tandukar, Gerald M Wilson
Hsp70 is a protein chaperone that prevents protein aggregation and aids protein folding by binding to hydrophobic peptide domains through a reversible mechanism directed by an ATPase cycle. However, Hsp70 also binds U-rich RNA including some AU-rich elements (AREs) that regulate the decay kinetics of select mRNAs and has recently been shown to bind and stabilize some ARE-containing transcripts in cells. Previous studies indicated that both the ATP- and peptide-binding domains of Hsp70 contributed to the stability of Hsp70-RNA complexes and that ATP might inhibit RNA recruitment...
August 25, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28671094/expression-of-mlaa34-hsp70-fusion-gene-constructed-by-soe-pcr
#15
Chen Zhao, Jiaqi Zhao, Wanying Wang, Yening Fan, Chunping Ma, Donghong Zhang, Yang Lv
To construct the pIRES2-MLAA34-HSP70 recombinant vector and express the MLAA34-HSP70 recombinant proteins in Escherichia coli (E. coli). The MLAA34 and the HSP70 genes were extracted from U937 cells by RT-PCR, and then we amplified the fusion gene MLAA34-HSP70 by SOE-PCR and inserted it into the pIRES2-EGFP vector to construct the pIRES2-MLAA34-HSP70 recombinant vector. We amplified the fusion gene MLAA34-HSP70 successfully and identified the correctness of pIRES2-MLAA34-HSP70 recombinant vector by PCR and restriction endonuclease...
May 2017: Pakistan Journal of Pharmaceutical Sciences
https://www.readbyqxmd.com/read/28666159/optimization-of-expression-and-purification-of-human-mortalin-hsp70-folding-unfolding-analysis
#16
Mohd Shahnawaz Khan, Anwar Ahmed, Shams Tabrez, Badar Ul Islam, Nayyar Rabbani, Ajamaluddin Malik, Mohamad A Ismael, Mohammad A Alsenaidy, Abdulrahman M Alsenaidy
Human mortalin is a Hsp70 mitochondrial protein that plays an essential role in the biogenesis of mitochondria. The deregulation of mortalin expression and its functions could lead to several age-associated disorders and some types of cancers. In the present study, we optimized the expression and purification of recombinant human mortalin by the use of two-step chromatography. Low temperature (18°C) and 0.5mM (IPTG) was required for optimum mortalin expression. Chaperone activity of mortalin was assessed by the citrate synthase and insulin protection assay, which suggested their protective role in mitochondria...
June 12, 2017: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
https://www.readbyqxmd.com/read/28627344/-protective-effect-of-bag-1l-mediated-by-lentivirus-in-human-neuroblastoma-cells-induced-by-hypoxia
#17
Yun Wang, Qingshu Li, Chao Jia, Chunyu Xie, Yankun Song, Nan Zhang, Yan Qu
OBJECTIVE: To investigate the protective effects of lentivirus mediated Bcl-2-associated athanogene 1L (BAG-1L) over-expression on human neuroblastoma cells (SH-SY5Y) induced by hypoxia/re-oxygenation, and to study its effect on the phosphoinositide 3 kinase serine/threonine protein kinase (PI3K/AKT) pathway. METHODS: SH-SY5Y cells were cultured in vitro, and the cells at logarithmic phase were collected, and they were divided into recombined lentiviral infection group [infected by lentivirus containing BAG-1L and green fluorescent protein (GFP) gene], vector control group (infected by lentivirus containing GFP without BAG-1L gene) and cell control group (non-infection)...
March 2017: Zhonghua Wei Zhong Bing Ji Jiu Yi Xue
https://www.readbyqxmd.com/read/28576342/development-of-a-colloidal-gold-immunochromatographic-strip-based-on-hsp70-for-the-rapid-detection-of-echinococcus-granulosus-in-sheep
#18
Xunhui Zhuo, Yingchao Yu, Xueqiu Chen, Zhuangzhi Zhang, Yi Yang, Aifang Du
Echinococcus granulosus is the causative pathogen of cystic echinococcosis, a serious disease endangering human and animal health. In this study, an immunochromatographic strip was developed based on the recombinant protein Heat shock protein 70 (HSP70) for the serological detection of E. granulosus. The protocol completes within 20min requiring no specialized equipment or chemical reagents, while specificity tests confirmed no cross-reactivity with positive serum of Fasciola hepatica, Haemonchus contortus, Peste des petits ruminants virus (PPRV) and Foot-and-mouth disease virus (FMDV)...
June 15, 2017: Veterinary Parasitology
https://www.readbyqxmd.com/read/28559404/peripheral-t-cell-reactivity-to-heat-shock-protein-70-and-its-cofactor-grpe-from-tropheryma-whipplei-is-reduced-in-patients-with-classical-whipple-s-disease
#19
Lucia Trotta, Kathleen Weigt, Katina Schinnerling, Anika Geelhaar-Karsch, Gerrit Oelkers, Federico Biagi, Gino Roberto Corazza, Kristina Allers, Thomas Schneider, Ulrike Erben, Verena Moos
Classical Whipple's disease (CWD) is characterized by the lack of specific Th1 response toward Tropheryma whipplei in genetically predisposed individuals. The cofactor GrpE of heat shock protein 70 (Hsp70) from T. whipplei was previously identified as a B-cell antigen. We tested the capacity of Hsp70 and GrpE to elicit specific proinflammatory T-cell responses. Peripheral mononuclear cells from CWD patients and healthy donors were stimulated with T. whipplei lysate or recombinant GrpE or Hsp70 before levels of CD40L, CD69, perforin, granzyme B, CD107a, and gamma interferon (IFN-γ) were determined in T cells by flow cytometry...
August 2017: Infection and Immunity
https://www.readbyqxmd.com/read/28447215/interplay-between-recombinant-hsp70-and-proteasomes-proteasome-activity-modulation-and-ubiquitin-independent-cleavage-of-hsp70
#20
Alexey V Morozov, Tatiana M Astakhova, David G Garbuz, George S Krasnov, Natalia V Bobkova, Olga G Zatsepina, Vadim L Karpov, Michail B Evgen'ev
The heat shock protein 70 (Hsp70, human HSPA1A) plays indispensable roles in cellular stress responses and protein quality control (PQC). In the framework of PQC, it cooperates with the ubiquitin-proteasome system (UPS) to clear damaged and dysfunctional proteins in the cell. Moreover, Hsp70 itself is rapidly degraded following the recovery from stress. It was demonstrated that its fast turnover is mediated via ubiquitination and subsequent degradation by the 26S proteasome. At the same time, the effect of Hsp70 on the functional state of proteasomes has been insufficiently investigated...
April 26, 2017: Cell Stress & Chaperones
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