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#32789395   2020/08/13 To Up

A simplified protein purification method through nickel cleavage of the recombinant protein from the Escherichia coli cell surface.

To simplify the protein purification process, we developed a novel one-step purification method in which the recombinant protein can be cleaved directly from the Escherichia coli cell surface. This method involves fusion of the target protein to the C-terminus of a LOS tag comprising a surface anchor protein (Lpp-OmpA) and a sequence-specific nickel-assisted cleavage (SNAC)-tag. The LOS tag facilitates the anchoring of the target protein to the outer membrane of E. coli cells and its separation from the cell membrane through Ni2+ cleavage. Intact, biologically active protein with a purity of 95% and a yield of approximately 100 mg per liter of culture can be readily obtained through Ni2+ cleavage in resuspension solution followed by centrifugation. In this study, a practical and promising protein purification method has been established with minimal labor and cost, as no cell disruption and chromatographic separation are required downstream.
Shanqing Huang, Tianbiao Wei, Wanxing Sha, Qingyuan Hu, Yingying Zhang, Jue Wang, Yufei Jiang, Hao Chen

1755 related Products with: A simplified protein purification method through nickel cleavage of the recombinant protein from the Escherichia coli cell surface.

200ul1010 100 U21mg250 10ìg1mg50

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#32788917   2020/08/03 To Up

Production of glycosylphosphatidylinositol-anchored proteins for vaccines and directed binding of immunoliposomes to specific cell types.

Liposomes are highly useful carriers for delivering drugs or antigens. The association of glycosylphosphatidylinositol (GPI)-anchored proteins to liposomes potentially enhances the immunogenic effect of vaccine antigens by increasing their surface concentration. Furthermore, the introduction of a universal immunoglobulin-binding domain can make liposomes targetable to virtually any desired receptor for which antibodies exist.
Wesley L Fotoran, Nicole Kleiber, Thomas Müntefering, Eva Liebau, Gerhard Wunderlich

1054 related Products with: Production of glycosylphosphatidylinositol-anchored proteins for vaccines and directed binding of immunoliposomes to specific cell types.

1050 ml1.00 flask510025 Tests1 kit10 ml1 kit100 ml

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#32788388   2020/08/11 To Up

eCLIPs bifurcation remodeling system for treatment of wide neck bifurcation aneurysms with extremely low dome-to-neck and aspect ratios: a multicenter experience.

Wide necked bifurcation aneurysms (WNBA) are among the most difficult aneurysms to treat. Very low dome-to-neck (DTN) and aspect ratios provide an even greater challenge in the management of WNBAs. We present the safety and efficacy profile for endovascular clip system (eCLIPs) device in the treatment of this subset of WNBAs with very unfavorable morphologies.
Joost De Vries, Hieronymus D Boogaarts, Leif Sørensen, Markus Holtmannspoetter, Goetz Benndorf, Bernd Turowski, Georg Bohner, Shahram Derakhshani, Chema Navasa, Wim H van Zwam, Michael Söderman, Riitta Rautio, Christian Mathys, Howard Riina, Thomas R Marotta

2025 related Products with: eCLIPs bifurcation remodeling system for treatment of wide neck bifurcation aneurysms with extremely low dome-to-neck and aspect ratios: a multicenter experience.



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#32788219   2020/08/11 To Up

The chemokine X-factor: structure-function analysis of the CXC motif at CXCR4 and ACKR3.

The human chemokine family consists of 46 protein ligands that induce chemotactic cell migration by activating a family of 23 G protein-coupled receptors. The two major chemokine subfamilies, CC and CXC, bind distinct receptor subsets. A sequence motif defining these families, the X position in the CXC motif, is not predicted to make significant contacts with the receptor, but instead links structural elements associated with binding  and activation. Here we use comparative analysis of chemokine NMR structures, structural modeling, and molecular dynamic simulations that suggested the X position reorients the chemokine N-terminus. Using CXCL12 as a model CXC chemokine, deletion of the X residue (P10) had little to no impact on the folded chemokine structure but diminished CXCR4 agonist activity as measured by ERK phosphorylation, chemotaxis, and Gi/o-mediated cAMP inhibition. Functional impairment was attributed to over 100-fold loss of CXCR4 binding affinity. Binding to the other CXCL12 receptor, ACKR3, was diminished  nearly 500-fold.  Deletion of P10 had little effect on CXCL12 binding to the CXCR4 N-terminus, a major component of the chemokine-GPCR interface. Replacement of the X residue with the most frequent amino acid at this position (P10Q) had an intermediate effect between wild-type and P10del in each assay, with ACKR3 having a higher tolerance for this mutation. This work shows that the X residue helps to position the CXCL12 N-terminus for optimal docking into the orthosteric pocket of CXCR4 and suggests that the CC/CXC motif contributes directly to receptor selectivity by orienting the chemokine N-terminus in a subfamily-specific direction.
Michael J Wedemeyer, Sarah A Mahn, Anthony E Getschman, Kyler S Crawford, Francis C Peterson, Adriano Marchese, John D McCorvy, Brian F Volkman

2904 related Products with: The chemokine X-factor: structure-function analysis of the CXC motif at CXCR4 and ACKR3.

0.1 mg5ug500 Units100.00 ug1100 100μg1mg100 1

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#32788216   2020/08/11 To Up

---A seven-residue deletion in PrP leads to generation of a spontaneous prion formed from C-terminal C1 fragment of PrP.

Prions result from a drastic conformational change of the host-encoded cellular prion protein (PrP), leading to the formation of beta-sheet-rich, insoluble and protease-resistant self-replicating assemblies (PrP). The cellular and molecular mechanisms involved in spontaneous prion formation in sporadic and inherited human prion diseases or equivalent animal diseases are poorly understood, in part because cell models of spontaneously-forming prions are currently lacking. Here, extending studies on the role of H2 alpha-helix C-terminus of PrP, we found that deletion of the highly conserved HTVTTTT segment of ovine PrP led to spontaneous prion formation in the RK13 rabbit kidney cell model. On long-term passage, the mutant cells stably produced proteinase-K resistant, insoluble and aggregated assemblies that were infectious for naïve cells expressing either the mutant protein or other PrPs with slightly different deletions in the same area. The electrophoretic pattern of PK-resistant core of the spontaneous prion (∆) contained mainly C-terminal polypeptides akin to C1, the cell-surface anchored C-terminal moiety of PrP generated by natural cellular processing. RK13 cells expressing solely ∆190-196 C1 PrP construct, in absence of the full-length protein, were susceptible to ∆ prions. ∆ infection induced the conversion of the mutated C1 into a PK-resistant and infectious form perpetuating the biochemical characteristics of ∆Spont prion. In conclusion this work provides a unique cell-derived system generating spontaneous prions and provides evidence that the 113 C-terminal residues of PrP are sufficient for a self-propagating prion entity.
Carola Munoz-Montesino, Djabir Larkem, Clément Barbereau, Angélique Igel-Egalon, Sandrine Truchet, Eric Jacquet, Naïma Nhiri, Mohammed Moudjou, Christina Sizun, Human Rezaei, Vincent Béringue, Michel Dron

2700 related Products with: ---A seven-residue deletion in PrP leads to generation of a spontaneous prion formed from C-terminal C1 fragment of PrP.

100ug Lyophilized1 kit96 tests0.5 mg0.5 mg100ug Lyophilized100 mg0.1mg100 μg100 μg100 μg

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#32787922   2020/08/12 To Up

Prominent microglial inclusions in transgenic mouse models of α-synucleinopathy that are distinct from neuronal lesions.

Alpha-synucleinopathies are a group of progressive neurodegenerative disorders, characterized by intracellular deposits of aggregated α-synuclein (αS). The clinical heterogeneity of these diseases is thought to be attributed to conformers (or strains) of αS but the contribution of inclusions in various cell types is unclear. The aim of the present work was to study αS conformers among different transgenic (TG) mouse models of α-synucleinopathies. To this end, four different TG mouse models were studied (Prnp-h[A53T]αS; Thy1-h[A53T]αS; Thy1-h[A30P]αS; Thy1-mαS) that overexpress human or murine αS and differed in their age-of-symptom onset and subsequent disease progression. Postmortem analysis of end-stage brains revealed robust neuronal αS pathology as evidenced by accumulation of αS serine 129 (p-αS) phosphorylation in the brainstem of all four TG mouse lines. Overall appearance of the pathology was similar and only modest differences were observed among additionally affected brain regions. To study αS conformers in these mice, we used pentameric formyl thiophene acetic acid (pFTAA), a fluorescent dye with amyloid conformation-dependent spectral properties. Unexpectedly, besides the neuronal αS pathology, we also found abundant pFTAA-positive inclusions in microglia of all four TG mouse lines. These microglial inclusions were also positive for Thioflavin S and showed immunoreactivity with antibodies recognizing the N-terminus of αS, but were largely p-αS-negative. In all four lines, spectral pFTAA analysis revealed conformational differences between microglia and neuronal inclusions but not among the different mouse models. Concomitant with neuronal lesions, microglial inclusions were already present at presymptomatic stages and could also be induced by seeded αS aggregation. Although nature and significance of microglial inclusions for human α-synucleinopathies remain to be clarified, the previously overlooked abundance of microglial inclusions in TG mouse models of α-synucleinopathy bears importance for mechanistic and preclinical-translational studies.
Gaye Tanriöver, Mehtap Bacioglu, Manuel Schweighauser, Jasmin Mahler, Bettina M Wegenast-Braun, Angelos Skodras, Ulrike Obermüller, Melanie Barth, Deborah Kronenberg-Versteeg, K Peter R Nilsson, Derya R Shimshek, Philipp J Kahle, Yvonne S Eisele, Mathias Jucker

2483 related Products with: Prominent microglial inclusions in transgenic mouse models of α-synucleinopathy that are distinct from neuronal lesions.

100 μg100 ul1 mg100ug Lyophilized200 4 Membranes/Box100 μg100 μg100 μg500 100 μg1 mg

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#32787254   2020/08/07 To Up

Heavy-Metal-Free Fischer-Tropsch Type Reaction: Sequential Homologation of Alkylborane Using a Combination of CO and Hydrides as Methylene Source.

Carbon homologation reactions occur within the well-known Fischer-Tropsch process, usually mediated by transition metal catalysts at high temperature. Here we report the low-temperature, heavy-metal-free homologation of a carbon chain using CO as a C-source showing for the first time that transition-metal catalysts are not required for Fischer-Tropsch-type reactivity. Reaction of an alkylborane in the presence of either LiHBEt or LiAlH resulted in multiple CO insertion/reduction events to afford elongated chains by more than two methylene (-CH-) units, affording aldehyde products upon oxidative aqueous workup. Theoretical and experimental mechanistic studies indicate that the boron terminus is responsible for CO incorporation as well as sequential hydride delivery leading to reduction of acylborane intermediates to alkylboranes.
Andreas Phanopoulos, Shrinwantu Pal, Takafumi Kawakami, Kyoko Nozaki

2051 related Products with: Heavy-Metal-Free Fischer-Tropsch Type Reaction: Sequential Homologation of Alkylborane Using a Combination of CO and Hydrides as Methylene Source.



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#32786518   2020/08/11 To Up

Engineering a Plant Viral Coat Protein for Hybrid Self-Assembly of CO-Capturing Catalytic Nanofilaments.

Plant virus-based nanoparticles are used as self-assembled protein scaffolds for the construction of enzyme nanocarriers. To date, one-pot production and coupling of both enzymes and scaffolds by genetic conjugation has been demonstrated only in plants. Herein, we report bacterial production and self-assembly of nanofilaments for CO capture. Filamentous virus-like particles (VLPs) were successfully formed by genetically fusing carbonic anhydrase (CA) from to the terminus of the coat protein (CP) of potato virus Y with a flexible linker. The instability of VLPs against proteolytic degradation was circumvented by the periplasmic export of the fusion protein. The truncated form of CP co-expressed by internal translation was crucial for the successful formation of long filamentous VLPs by alleviating steric hindrance hybrid assembly. The fast and economic bottom-up fabrication of highly active nanobiocatalyst allows the nanofilaments to be efficiently used and recovered in potential biocatalytic and biosensor systems.
Suhan Wi, In Seong Hwang, Byung Hoon Jo

1083 related Products with: Engineering a Plant Viral Coat Protein for Hybrid Self-Assembly of CO-Capturing Catalytic Nanofilaments.

0.1 mg10000.1 mg1mg10ìg500100μg100ug0.2 mg1000.25 mg100ul

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#32786352   2020/08/05 To Up

Optogenetic control of heterologous metabolism in E. coli.

Multi-objective optimization of microbial chassis for the production of xenobiotic compounds requires the implementation of metabolic control strategies that permit dynamic distribution of cellular resources between biomass and product formation. We addressed this need in a previous study by engineering the T7 RNA polymerase to be thermally responsive. The modified polymerase is activated only after the temperature of the host cell falls below 18oC, and Escherichia coli cells that employ the protein to transcribe the heterologous lycopene biosynthetic pathway exhibit impressive improvements in productivity. We have expanded our toolbox of metabolic switches in the current study by engineering a version of the T7 RNA polymerase that drives the transition between biomass and product formation upon stimulation with red light. The engineered polymerase is expressed as two distinct polypeptide chains. Each chain comprises one of two photoactive components from Arabidopsis thaliana, phytochrome B (PhyB) and phytochrome-integrating factor 3 (PIF3), as well as the N- or C-terminus domains of both, the vacuolar ATPase subunit (VMA) intein of Saccharomyces cerevisiae and the polymerase. Red light drives photodimerization of PhyB and PIF3, which then brings together the N- and C-terminus domains of the VMA intein. Trans-splicing of the intein follows suit and produces an active form of the polymerase that subsequently transcribes any sequence that is under the control of a T7 promoter. The photodimerization also involves a third element, the cyanobacterial chromophore phycocyanobilin (PCB), which too is expressed heterologously by E. coli. We deployed this version of the T7 RNA polymerase to control the production of lycopene in E. coli and observed tight control of pathway expression. We tested a variety of expression configurations to identify one that imposes the lowest metabolic burden on the strain, and we subsequently optimized key parameters such as the source, moment and duration of photostimulation. We also identified targets for future refinement of the circuit. In summary, our work is a significant advance for the field and greatly expands on previous work by other groups that have used optogenetic circuits to control heterologous metabolism in prokaryotic hosts.
Adhithi Ravi Raghavan, Kevin Salim, Vikramaditya G Yadav

2632 related Products with: Optogenetic control of heterologous metabolism in E. coli.

96 tests-

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