Topics

Expression, purification, and efficient refolding of the extracellular domain of Escherichia coli-expressed signaling receptor herpesvirus entry mediator.

08:00 EDT 9th July 2019 | BioPortfolio

Summary of "Expression, purification, and efficient refolding of the extracellular domain of Escherichia coli-expressed signaling receptor herpesvirus entry mediator."

Herpesvirus entry mediator (HVEM), a member of the TNF-receptor superfamily, plays an important role in the regulation of the immune system. It forms a complex with ligands and can either activate or inhibit the response of the immune system. Furthermore, HVEM can exhibit pro-inflammatory or anti-inflammatory effects in many human diseases. Therefore, understanding the mechanism underlying the interaction of HVEM with other receptors is extremely important to design small therapeutic molecules that can stimulate the response of the immune system. In this study, we attempted to develop the most efficient method for the expression and purification of the extracellular domain of HVEM using Escherichia coli. The soluble fraction constituted only a small portion of the E. coli-expressed protein, whereas most majority of the protein was found to be accumulated in the insoluble fraction. Three different protein refolding methods were analyzed: dialysis, dilution, and using chromatographic column. The oligomeric state of the protein was determined by characterizing the obtained fractions using analytical size exclusion chromatography. All the obtained fractions were tested for their ability to form a complex with B- and T-lymphocyte attenuator using enzyme-linked immunosorbent assay. The results of this study provide crucial information regarding the production of HVEM protein in a robust, well-established, and convenient heterologous expression system using E. coli as a host. In addition, it allows for the selection of the most effective method for appropriate refolding of HVEM protein, which gets accumulated in the insoluble fraction.

Affiliation

Journal Details

This article was published in the following journal.

Name: Protein expression and purification
ISSN: 1096-0279
Pages: 105450

Links

DeepDyve research library

PubMed Articles [21207 Associated PubMed Articles listed on BioPortfolio]

The NT11, a novel fusion tag for enhancing protein expression in Escherichia coli.

The Escherichia coli (E. coli) expression system has been widely used to produce recombinant proteins. However, in some heterologous expressions, there are still difficulties in large-scale production...

Expression and Purification of a Functional E. coli CH-Methionine-Labeled Thermostable Neurotensin Receptor 1 Variant for Solution NMR Studies.

Escherichia coli (E. coli) is the most widely used expression host for recombinant proteins due to high expression yields and straightforward molecular cloning. Directed evolution of G protein-coupled...

Recombinant expression, purification and characterization of acetylated LysargiNase from Escherichia coli with high activity and stability.

LysargiNase is a novel characterized metalloprotease that can cleave N-terminal of lysine or arginine residue. The peptides generated by LysargiNase were just mirrors to those by trypsin. This charact...

Soluble expression of mature Rhizopus chinensis lipase in Escherichia coli and enhancement of its ester synthesis activity.

The production of membrane-associated lipase from Rhizopus chinensis (RCL), which has a high ester synthesis activity and important potential applications, is difficult in heterologous expression syst...

Enhancement of the stability of Mycobacterium tuberculosis recombinant antigen expressed in Escherichia coli using cell lysis additives.

Mycobacteria tuberculosis (Mtb), the causative agent of tuberculosis, is a slow-growing bacterium. Expression in Escherichia coli is a widely used method for large-scale production of diagnostic antig...

Clinical Trials [3026 Associated Clinical Trials listed on BioPortfolio]

A Clinical Trial to Evaluate a Recombinant Staphylococcus Aureus Vaccine (Escherichia Coli) in Healthy Adults

This is a single center, open-label phase1b clinical trial. The study will evaluate the safety and immunogenicity of an experimental recombinant staphylococcus aureus vaccine with differen...

Enteropathogenic Escherichia Coli (EPEC): Does it Have a Role in Colorectal Tumourigenesis?

Despite the characterization of many aetiologic genetic changes. The specific causative factors in the development of sporadic colorectal cancer remain unclear. This study was performed to...

Epidemiology of ST131 in Besançon University Hospital

The sequence type 131 (ST131) is a predominant lineage among extraintestinal pathogenic Escherichia coli. It plays a major role in the worldwide dissemination of E. coli that produce exten...

A Phase Ⅳ Clinical Trial of the Recombinant Hepatitis E Vaccine (Escherichia Coli)(the Lot Consistency Trial)

This study is to evaluate lot-lot consistency of Recombinant Hepatitis E Vaccine (Escherichia Coli) Hecolin®.

Characteristics of Lower Respiratory Tract Escherichia Coli Isolates in Mechanically Ventilated Intensive Care Patients

Prospective, multicenter observational study to collect Escherichia coli (E. coli) isolates originating from mechanically ventilated intensive care unit (ICU) patients; in order to charact...

Medical and Biotech [MESH] Definitions

Strains of ESCHERICHIA COLI that are a subgroup of SHIGA-TOXIGENIC ESCHERICHIA COLI. They cause non-bloody and bloody DIARRHEA; HEMOLYTIC UREMIC SYNDROME; and hemorrhagic COLITIS. An important member of this subgroup is ESCHERICHIA COLI O157-H7.

Strains of Escherichia coli that possess virulence traits which allow them to invade, colonize, and induce disease in tissues outside of the GASTROINTESTINAL TRACT. They are a cause of URINARY TRACT INFECTIONS (UROPATHOGENIC ESCHERICHIA COLI); neonatal MENINGITIS; SEPSIS; PNEUMONIA; and SURGICAL WOUND INFECTION.

An enterohemorrhagic Escherichia coli of the O subfamily that can cause severe FOODBORNE DISEASE. The H4 serotype strain produces SHIGA TOXINS and has been linked to human disease outbreaks, including some cases of HEMOLYTIC-UREMIC SYNDROME, resulting from contamination of foods by feces containing E. coli O104.

A species of gram-negative, rod-shaped bacteria belonging to the K serogroup of ESCHERICHIA COLI. It lives as a harmless inhabitant of the human LARGE INTESTINE and is widely used in medical and GENETIC RESEARCH.

A verocytotoxin-producing serogroup belonging to the O subfamily of Escherichia coli which has been shown to cause severe food-borne disease. A strain from this serogroup, serotype H7, which produces SHIGA TOXINS, has been linked to human disease outbreaks resulting from contamination of foods by E. coli O157 from bovine origin.

Quick Search


DeepDyve research library

Relevant Topic

Cytokine Tumour Necrosis Factor (TNF)
TNF is a compound that is classified as a cytokine which plays a central role in the cellular mechanisms of apoptosis or cell death. However, there are a number of different kinds of TNF, just under twenty, but the family of molecules have very similar a...


Searches Linking to this Article