Expression, purification, crystallization and preliminary X-ray crystallographic analysis of fructose-1,6-bisphosphate aldolase from Escherichia coli
Fructose-1,6-bisphosphate aldolase is one of the most important enzymes in the glycolytic pathway and catalyzes the reversible cleavage of fructose-1,6-bisphosphate to dihydroxyacetone phosphate and glyceraldehyde 3-phosphate. The full-length fbaB gene encoding fructose-1,6-bisphosphate aldolase class I (FBPA I) was cloned from Escherichia coli strain BL21. FBPA I was overexpressed in E. coli and purified. Biochemical analysis found that the optimum reaction temperature of FBPA I is 330.5 K and that the enzyme has a high temperature tolerance. Crystals of recombinant FBPA I were obtained by the sitting-drop vapour-diffusion technique in a condition consisting of 19 mg ml−1 FBPA I in 0.1 M Tris pH 9.0, 10%(w/v) polyethylene glycol 8000 and diffracted to 2.0 Å resolution. The crystals belonged to the monoclinic space group C2, with unit-cell parameters a = 217.7, b = 114.9, c = 183.9 Å, β = 124.6°. The asymmetric unit of these crystals may contain ten molecules, giving a Matthews coefficient of 2.48 Å3 Da−1 and a solvent content of 50.5%.
Publication date: Available online 1 April 2020Source: Food ControlAuthor(s): Marie Frétin, Christophe Chassard, Céline Delbès, René Lavigne, Etienne Rifa, Sébastien Theil, Benoit Fernandez, Patrice Laforce, Cécile Callon
CONCLUSION: These results suggested that rHP-NAP possesses the potential for use as an adjuvant of dendritic cell-based vaccine in anti-melanoma treatment. PMID: 32224538 [PubMed - in process]
Conclusion: To facilitate transparent consent, single-centre rather than pooled outcome data should be utilized. PMID: 32223494 [PubMed - as supplied by publisher]
Contributors : Subhash C Verma ; Sankar Adhya ; Soumya G Remesh ; Michal HammelSeries Type : Expression profiling by high throughput sequencingOrganism : Escherichia coliNucleoid remodeling facilitated by DNA supercoiling results in changes in nucleoid configuration and involves nucleoid-associated proteins (NAPs) and structural maintenance of chromosomes (SMC) proteins among others. Changes in nucleoid configurations regulated by NAPs are synchronized with cellular adaptation and influence the simultaneous expression of several genes. HU, a ubiquitous bacterial histone-like protein, is among the most conserved and abunda...
We report the synthesis and expression measurement of ~9000 inducible promoter variants. By tagging individual variants with barcodes, we can measure the expression levels of all variants under both induced and uninduced conditions in a single pooled experiment. Sequencing data here is used for 1) paired-end sequencing to map barcodes to their promoters or 2) Quantifying the counts of barcodes at the DNA and RNA levels
Contributors : Pabitra Nandy ; Savita Chib ; Aswin S SeshasayeeSeries Type : Expression profiling by high throughput sequencingOrganism : Escherichia coliRNA Sequencing of wild type Eschrichia coli and a RNA polymerase mutant isolated from deep stationary phase
Contributors : Fernando H Martins ; Ashwani Kumar ; Chao Xing ; Vanessa Sperandio ; Waldir P EliasSeries Type : Expression profiling by high throughput sequencingOrganism : Homo sapiensAtypical enteropathogenic Escherichia coli (aEPEC) is amongst the leading causes of diarrheal disease worldwide. The colonization of the gut mucosa by aEPEC results in actin pedestal formation at the site of bacterial attachment. This cytoskeletal rearrangement is triggered by the interaction between the bacterial adhesin intimin and its receptor Tir, which is translocated through the type three secretion system, to the host cell. While some...
by Porntida Kobpornchai, Robin J. Flynn, Onrapak Reamtong, Nonglucksanawan Rittisoonthorn, Nathamon Kosoltanapiwat, Kobporn Boonnak, Usa Boonyuen, Sumate Ampawong, Montakan Jiratanh, Muncharee Tattiyapong, Poom AdisakwattanaTrichinella spiralis can modulate host immune responses to retain a suitable environment for its long-term survival. Incidentally, the parasite elicits regulatory effects through immunomodulatory molecule release, which can suppress host inflammation and may be used for the treatment of unrelated inflammatory diseases in someday. Here we identified and characterized a novelT.spiralis cystatin (TsCstN), ...
Publication date: Available online 30 March 2020Source: Microbial PathogenesisAuthor(s): Jennifer Anne Pienaar, Atheesha Singh, Tobias George Barnard
The Journal of Physical Chemistry BDOI: 10.1021/acs.jpcb.0c01595