Only in Titles

Search results for: Recombinant E. coli Hsp60 GroEL Proteins

paperclip

#29253594   2017/12/16 To Up

Evaluation of immune responses induced by polymeric OMP25-BLS Brucella antigen.

Brucellosis is one the serious infectious diseases caused deleterious health and economic losses. Vaccination with subunit vaccines is the efficient alternative way than live attenuated vaccines against infectious diseases. Herein a new chimeric OMP25-BLS antigen emulsified in Chitosan Nanoparticles was designed and its immune responses were compared with control groups. Also, the role of heat shock protein 60 kDa in combination with OMP25-BLS antigen was assessed. Structural and antigenic features of chimeric antigen were predicted using bioinformatics tools. Moreover, the humoral and cellular immune responses were measured by ELISA in seven different groups. Observations showed rOMP25-BLS structure was highly stable and antigenic. Cytokines analysis showed rOMP25 and rOMP25-BLS + rHSP60 induced higher titer of INF-γ than rHSP60 and rOMP25-BLS. There was no statistically significant difference between positive control group and rOMP25-BLS + rHSP60 in inducing TNF-α (p < .05). Additionally, the highest titer of IL-4 was dedicated to rOMP25 among other immunized treatments, while there were no significant differences between positive control group and other immunized groups with recombinant proteins (p < .05). In addition, rOMP25-BLS and rHSP60 induced higher titer of total antibody compared to other groups. Also, rHSP60 could improve IgG2a to IgG1 ratio when it used in combination with chimeric antigen. Moreover, the lymphocyte proliferation index was higher in chimeric rOMP25-BLS + HSP60 antigen. In conclusion, while rOMP25-BLS chimeric antigen unable to induce efficient cellular response than individual injection of rOMP25, its injection in combination with rHSP60 could improve cellular immunity.
Soheil Yousefi, Tooba Abbassi-Daloii, Mohammad Hadi Sekhavati, Mojtaba Tahmoorespur

1306 related Products with: Evaluation of immune responses induced by polymeric OMP25-BLS Brucella antigen.

25 100 µg50ul1 mg100 µg0.1ml0.1ml (1.3mg/ml)1mg100 µg96T50ul1 kit

Related Pathways

paperclip

#29037137   // To Up

Heterologous Expression of Chaperones from Hyperthermophilic Archaea Inhibits Aminoglycoside-Induced Protein Misfolding in Escherichia coli.

Aminoglycoside antibiotics affect protein translation fidelity and lead to protein aggregation and an increase in intracellular oxidative stress level as well. The overexpression of the chaperonin GroEL/GroES system promotes short-term tolerance to aminoglycosides in Escherichia coli. Here, we demonstrated that the coexpression of prefoldin or Hsp60 originating from the hyperthermophilic archaeon Pyrococcus furiosus in E. coli cells can rescue cell growth and inhibit protein aggregation induced by streptomycin exposure. The results of our study show that hyperthermophilic chaperones endow E. coli with a higher tolerance to streptomycin than the GroEL/GroES system, and that they exert better effects on the reduction of intracellular protein misfolding, indicating that these chaperones have unique features and functions.
S Peng, Z Chu, J Lu, D Li, Y Wang, S Yang, Y Zhang

1631 related Products with: Heterologous Expression of Chaperones from Hyperthermophilic Archaea Inhibits Aminoglycoside-Induced Protein Misfolding in Escherichia coli.

50 2x 100ug100 10 96T1 Set1 mL10 1 Set1 Set1 Set50

Related Pathways

paperclip

#23643628   2013/04/30 To Up

Chaperonin GroEL: a novel phylogenetically conserved protein with strong immunoreactivity of Avian Pathogenic Escherichia coli isolates from duck identified by immunoproteomics.

Avian Pathogenic Escherichia coli (APEC) is one of the most important bacterial pathogens of poultry. The lack of suitable vaccines and the emergence of multi-resistant strains have hampered the control of avian colibacillosis. To identify immunogenic proteins of APEC as vaccine candidates, immunoproteomics and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) were applied. Proteins from total cell lysates of APEC DE205B isolated from the brain of a duck with septicemia and neurological symptom in China were separated by two-dimensional electrophoresis (2-DE) and reacted with hyperimmune duck serum against DE205B. Fourteen immunoreactive spots were found, representing 11 distinct proteins. These included two predominant immunogenic components, outer membrane protein A (OmpA) and flagellin (FliC). GroEL, which is a member of the molecular chaperone family and identical structurally to eukaryotic heat shock protein 60 (Hsp60), and the other eight antigens are reported here as immunoreactive proteins of APEC for the first time. Subsequently, nine genes encoding the identified proteins were successfully cloned and expressed in E. coli BL21 (DE3). Seven of the recombinant proteins were able to react with hyperimmune duck serum and three of them, GroEL, OmpA and FliC, showed stronger immunoreactivity. Challenge studies revealed that, just like OmpA and FliC, recombinant GroEL stimulated a strong antibody response and supported protective efficacy against APEC infection in ducks. With high phylogenetic conservation, it is considered that GroEL would be an ideal immunogen of APEC for vaccine development.
Yinli Bao, Zhipeng Zhai, Shaohui Wang, Jiale Ma, Wei Zhang, Chengping Lu

1847 related Products with: Chaperonin GroEL: a novel phylogenetically conserved protein with strong immunoreactivity of Avian Pathogenic Escherichia coli isolates from duck identified by immunoproteomics.

100ug100ug50 10 10ìg1mg50 200 1mg100ug200 100

Related Pathways

paperclip

#23474314   2013/03/06 To Up

Recombinant heat shock protein 60 (Hsp60/GroEL) of Salmonella enterica serovar Typhi elicits cross-protection against multiple bacterial pathogens in mice.

Heat shock proteins (HSPs) or stress proteins are recognized as protective antigens against a wide range of bacterial diseases. Conservation of HSPs across different life forms also appears to contribute to the antigenicity of these proteins. Due to their high sequence homology, there exists an immunological cross-recognition between different bacterial species. In the present study, we evaluated the efficacy of recombinant GroEL of Salmonella enterica serovar Typhi as a vaccine candidate against various bacterial pathogens viz.; Shigella dysenteriae type I, Shigella flexneri, Shigella boydii, enteropathogenic Escherichia coli (EPEC), Klebsiella pneumoniae and Pseudomonas aeruginosa. In vitro serum bactericidal assay (SBA) with GroEL antisera showed 50-55% inhibition of cells of Shigella Spp., 65-75% of E. coli, 60-65% of K. pneumoniae, 45-50% of P. aeruginosa. In in vivo experiments, mice immunized with GroEL protein of S. Typhi showed 60-65% protection against S. flexneri, S. dysenteriae type I, S. boydii. Similarly 75-80% protection was observed against enteropathogenic E. coli, 70-80% against K. pneumoniae. 50% of mice survived the lethal infection against P. aeruginosa. Organ burden and histopathological studies also revealed significant reduction of bacterial infection. This study shows the cross-protective efficacy of recombinant GroEL of S. Typhi which could lead to the development of a single vaccine candidate protective against multiple bacterial pathogens.
S T S Chitradevi, G Kaur, K Singh, R Sugadev, A Bansal

1402 related Products with: Recombinant heat shock protein 60 (Hsp60/GroEL) of Salmonella enterica serovar Typhi elicits cross-protection against multiple bacterial pathogens in mice.

100 μg101mg10 ìg25 μg1mg2010 μg2050.1 mg10 μg

Related Pathways

paperclip

#22890156   2012/08/05 To Up

Heat shock protein 60 of filarial parasite Brugia malayi: cDNA cloning, expression, purification and in silico modeling and analysis of its ATP binding site.

We report here cloning and expression of full length mitochondrial HSP60 gene of Brugia malayi adult worm (mtHSP60bm), purification of the gene product by affinity chromatography, its in silico 3D structure and the sequence homology of the protein with Escherichia coli GroEL/ES and human HSP60. The ATP binding pocket of human HSP60 and mtHSP60bm were analyzed and compared using in silico models. The distribution of HSP60 in different life-stages of the parasite was determined using antibodies raised against recombinant mtHSP60bm (rmtHSP60bm). mtHSP60bm was present in all life-stages of the parasite except third stage infective larvae, in which it could be induced by heat-shock, and showed high degree of homology with E. coli GroEL/ES. The ATP binding pocket of HSP60 in humans, E. coli and B. malayi were also found structurally conserved. This similarity between human and mtHSP60bm might be useful in understanding the host-parasite interactions. This is the first ever report on distribution, cloning, sequence homology and ATP binding site of mtHSP60bm.
R C Misra, A K Verma, S K Verma, V Kumar, W A Siddiqui, M I Siddiqi, P K Murthy

2586 related Products with: Heat shock protein 60 of filarial parasite Brugia malayi: cDNA cloning, expression, purification and in silico modeling and analysis of its ATP binding site.

1000 TESTS/0.65ml10 ìg100.00 ug100 μg100 μg100 ul96T0.1 mg500 25 μg100.00 ug96T

Related Pathways

paperclip

#22160895   // To Up

Heat-shock protein fusion vectors for improved expression of soluble recombinant proteins in Escherichia coli.

Molecular chaperones or heat-shock proteins (HSPs) are protein machines that interact with unfolded or partially folded polypeptides and assist them in attaining their proper conformation. The folding reaction relies on a complex array of scaffolding effects and ATP-driven conformational changes that mediate the temporary unfolding and subsequent refolding of protein substrates. DnaK and GroEL are the two major Escherichia coli chaperones. They belong to the HSP70 and HSP60 families of proteins, respectively, and play a major role in protein folding. Here, we describe a set of bacterial expression vectors that permits the fusion of a protein of interest to DnaK or GroEL and its subsequent quantitative expression in a soluble, easily purifiable form. We also provide a set of compatible co-chaperone expression constructs that permit the simultaneous co-expression of the DnaK and GroEL physiological partners to further increase protein solubility. The system was successfully tested using the murine prion protein (PrP). Although PrP is normally insoluble when expressed in E. coli, we show that utilizing our vectors it can be produced in a soluble form as a DnaK or GroEL fusion. This system is useful for the production of a large array of proteins that fail to fold properly when expressed in E. coli.
Christos A Kyratsous, Christos A Panagiotidis

2440 related Products with: Heat-shock protein fusion vectors for improved expression of soluble recombinant proteins in Escherichia coli.

0.1 mg10 0.1mg50 100 100 100 100510 100 50

Related Pathways

paperclip

#20633027   // To Up

Mycobacterium tuberculosis employs Cpn60.2 as an adhesin that binds CD43 on the macrophage surface.

CD43 is a large sialylated glycoprotein found on the surface of haematopoietic cells and has been previously shown to be necessary for efficient macrophage binding and immunological responsiveness to Mycobacterium tuberculosis. Using capsular material from M. tuberculosis and recombinant CD43-Fc, we have employed affinity chromatography to show that Cpn60.2 (Hsp65, GroEL), and to a lesser extent DnaK (Hsp70), bind to CD43. Competitive inhibition using recombinant protein and polyclonal F(ab')(2) antibody-mediated epitope masking studies were used to evaluate M. tuberculosis binding to CD43(+/+) versus CD43(-/-) macrophages. Results showed that Cpn60.2, but not DnaK, acts as a CD43-dependent mycobacterial adhesin for macrophage binding. Assessment of the specific binding between Cpn60.2 and CD43 showed it to be saturable, with a comparatively weak affinity in the low micromolar range. We have also shown that the ability of Cpn60.2 to competitively inhibit M. tuberculosis binding to macrophages is shared by the Escherichia coli homologue, GroEL, but not by the mouse and human Hsp60 homologues. These findings add to a growing field of research that implicates molecular chaperones as having extracellular functions, including bacterial adherence to host cells. Thus, CD43 may act as a Pattern Recognition Receptor (PRR) for bacterial homologues of the 60 kDa molecular chaperone.
Tyler B M Hickey, Hermann J Ziltener, David P Speert, Richard W Stokes

2954 related Products with: Mycobacterium tuberculosis employs Cpn60.2 as an adhesin that binds CD43 on the macrophage surface.

200ul200ul200ul200ul0.025 mg200ul200ul0.025 mg0.25 25 G200ul

Related Pathways

paperclip

#19484809   // To Up

Immunogenicity and protective efficacy of GroEL (hsp60) of Streptococcus pneumoniae against lethal infection in mice.

The present study was carried out to evaluate the immunogenicity and protective efficacy of GroEL (hsp60) of Streptococcus pneumoniae, by expressing full length GroEL in heterologous host Escherichia coli BL21(DE3). PCR-amplified groEL was ligated in pQE 30 expression vector and subsequently transformed in E. coli DH5alpha strains. Cloning of groEL was confirmed by double digestion, followed by DNA sequencing. The His-tag containing recombinant GroEL was purified by Ni-NTA affinity chromatography. To determine the immunogenicity of GroEL, the mice were immunized by injecting 40 microg GroEL protein per mouse intraperitoneally. The results showed a significant increase in antibody titre and lymphocyte proliferation in animals immunized with GroEL as compared with control. Further, there was an appreciable increase in interleukin-2 (IL-2) and IL-4 production in lymphocytes isolated from immunized mice as compared with control. To determine the efficacy of GroEL in eliciting protection, the mice were challenged with the lethal dose of S. pneumoniae A66 type 3 capsular strain intranasally after the seventh day of the last immunization. In the GroEL-immunized mice the onset of death was insignificantly delayed and all the mice died by the seventh day postinfection.
Mohammed Nadeem Khan, Dhananjay Shukla, Anju Bansal, Sairam Mustoori, Govindaswami Ilavazhagan

1328 related Products with: Immunogenicity and protective efficacy of GroEL (hsp60) of Streptococcus pneumoniae against lethal infection in mice.

1mg20 5 G2055100ug1mg5096 tests

Related Pathways

paperclip

#19127371   2009/01/07 To Up

Conserved inserts in the Hsp60 (GroEL) and Hsp70 (DnaK) proteins are essential for cellular growth.

The Hsp60 and Hsp70 chaperones contain a number of conserved inserts that are restricted to particular phyla of bacteria. A one aa insert in the E. coli GroEL and a 21-23 insert in the DnaK proteins are specific for most Gram-negative bacteria. Two other inserts in DnaK are limited to certain groups of proteobacteria. The requirement of these inserts for cellular growth was examined by carrying out complementation studies with temperature-sensitive (T(s)) mutants of E. coli groEL or dnaK. Our results demonstrate that deletion or most changes in these inserts completely abolished the complementation ability of the mutant proteins. Studies with GroEL and DnaK from some other species that either lacked or contained these inserts also indicated that these inserts are essential for growth of E. coli. The DnaK from some bacteria contains a two aa insert that is not found in E. coli. Introduction of this insert into the E. coli DnaK also led to its inactivation, indicating that these inserts are specific for different groups. We postulate that these conserved inserts that are localized in loop regions on protein surfaces, are involved in some ancillary functions that are essential for the groups of bacteria where they are found.
Bhag Singh, Radhey S Gupta

1115 related Products with: Conserved inserts in the Hsp60 (GroEL) and Hsp70 (DnaK) proteins are essential for cellular growth.

10101001mg1mg1001mg10020205020

Related Pathways

paperclip

#18024219   2007/11/19 To Up

Cloning, expression and characterization of heat shock protein 60 (groEL) of Salmonella enterica serovar Typhi and its role in protective immunity against lethal Salmonella infection in mice.

Heat shock proteins (Hsps) represent dominant antigens in numerous microbial infections, suggesting a potential use of pathogen-derived Hsps for vaccination. The present study evaluates the immunogenicity and protective efficacy of groEL (Hsp60) of Salmonella enterica serovar Typhi against lethal challenge by S. Typhi Ty2 and Salmonella enterica serovar Typhimurium in mice. The groEL gene was cloned and expressed in Escherichia coli BL21 and purified by affinity chromatography. Immunization of mice with groEL resulted in a significant increase in antibody titers. Antibody isotyping revealed that groEL immunization induces both IgG1 and IgG2a antibodies. There was a significant increase in lymphocyte proliferation, interleukin-4 and interferon-gamma levels in cells isolated from immunized mice as compared to control. Immunization of mice with recombinant groEL protein with or without adjuvant conferred 70-90% protection against lethal infections either by S. Typhi Ty2 or S. Typhimurium. Passive immunization with anti-groEL sera also protected 50% mice against lethal infection.
Piyush Kumar Paliwal, Anju Bansal, Sarada S K Sagi, Sairam Mustoori, Ilavazhagan Govindaswamy

1677 related Products with: Cloning, expression and characterization of heat shock protein 60 (groEL) of Salmonella enterica serovar Typhi and its role in protective immunity against lethal Salmonella infection in mice.

0.1ml1 mg1 Set1 Set1 Set100ug Lyophilized1 Set1 Set1 Set100ug Lyophilized1 Set1 Set

Related Pathways