exploring the activity of tobacco etch virus protease in detergent solutions

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SelecTEV™ プロテアーゼ | 融合タンパク質のタグの切断・除去に | …- exploring the activity of tobacco etch virus protease in detergent solutions ,Exploring the activity of tobacco etch virus protease in detergent solutions. Analytical Biochemistry. 382: 69-71. Mohanty, A.K, Simmons, C.R, Wiener, M. (2002). Inhibition of tobacco etch virus protease activity by detergents. Protein Expression and Purification. 27: 109-114. ... Structural basis for the substrate specificity of tobacco etch ...Exploring the activity of tobacco etch virus protease in ...Tobacco etch virus (TEV) protease is generally used to remove affinity tags from target proteins. It has been reported that some detergents inhibit the activity of this protease, and therefore should be avoided when removing affinity tags from membrane proteins. The aim of this study was to explore and evaluate this further.



Combination of the mutations for improving activity of TEV ...

May 25, 2021·Tobacco etch virus protease (TEVp) is an enzymatic reagent to remove fusion tag, but additional purification steps are required for removing the TEVp after cleavage reaction is finished. Use of carrier-free and dependent TEVp immobilizates can eliminate protease contamination. In this work, we identified that, among the four constructed missense variants, the insoluble variant with the highest ...

Functional reconstitution of β2-adrenergic receptors ...

May 21, 2018·The percentage of active receptor in the detergent-solubilized β 2 AR stock, as determined by antagonist B max and A 280, was calculated at 97%. Approximately 90% of the binding activity for the radioligand, 3 H-DHA, was recovered on the Bio-Bead detergent removal step. After the size exclusion chromatography step, the receptor Nanodisc pool ...

Protocols and Video Articles Authored by Hans Hebert

Hans Hebert is the author of this article in the Journal of Visualized Experiments: "Method to Visualize and Analyze Membrane Interacting Proteins by Transmission Electron Microscopy"

Exploring the activity of tobacco etch virus protease in ...

Nov 01, 2008·Abstract. Tobacco etch virus (TEV) protease is generally used to remove affinity tags from target proteins. It has been reported that some detergents inhibit the activity of this protease, and therefore should be avoided when removing affinity tags from membrane proteins. The aim of this study was to explore and evaluate this further.

urn:nbn:se:kth:diva-18191 : Exploring the activity of ...

Tobacco etch virus (TEV) protease is generally used to remove affinity tags from target proteins. It has been reported that some detergents inhibit the activity of this protease, and therefore should be avoided when removing affinity tags from membrane proteins. The aim of this study was to explore and evaluate this further.

Razor(TM) TEV Protease | Biomol | Biomol

The TEV protease is a highly site-specific cysteine protease that is found in the Tobacco Etch Virus (TEV) often used for easy tag removal. One unit of Biomol Razor™ TEV Protease cleaves >85% of 3 µg of control substrate in 1 hour at 30°C. Advantage of Biomol Razor™ TEV Protease

Mechanism of hERG inhibition by gating-modifier toxin ...

Jan 07, 2021·Human ether-à-go-go-related gene potassium channel 1 (hERG) is a voltage-gated potassium channel, the voltage-sensing domain (VSD) of which is targeted by a gating-modifier toxin, APETx1. APETx1 is a 42-residue peptide toxin of sea anemone Anthopleura elegantissima and inhibits hERG by stabilizing the resting state. A previous study that conducted cysteine-scanning analysis of …

Disease-related mutations in PI3Kγ disrupt regulatory C ...

Mar 04, 2021·Cells were harvested after 55 hr of infection and frozen as described above. The frozen cell pellet was resuspended in lysis buffer (50 mM HEPES pH 7.5, 100 mM NaCl, 10 mM βME and protease inhibitor (Protease Inhibitor Cocktail Set III, Sigma)) and sonicated on ice for 1 min 30 s (15 s ON, 15 s OFF, power level 4.0) on the Misonix sonicator 3000.

SelecTEV™ プロテアーゼ | 融合タンパク質のタグの切断・除去に | …

Exploring the activity of tobacco etch virus protease in detergent solutions. Analytical Biochemistry. 382: 69-71. Mohanty, A.K, Simmons, C.R, Wiener, M. (2002). Inhibition of tobacco etch virus protease activity by detergents. Protein Expression and Purification. 27: 109-114. ... Structural basis for the substrate specificity of tobacco etch ...

Promoting Tag Removal of a MBP-Fused Integral Membrane ...

Sep 30, 2016·Exploring the activity of tobacco etch virus protease in detergent solutions. Analytical Biochemistry, 382 , 69–71. Article Google Scholar

LipaseMaturationFactorLMF1,MembraneTopologyand ...

binding peptide domain and a tobacco etch virus (TEV) prote-ase site and ending in two adjacent IgG-binding domains derived from protein A (18). All mouse and human LMF1 cDNA sequences, except for green fluorescent protein (GFP) fusions, were subcloned into the pcDNA3.1 expression vector (Invitrogen) containing an amino-terminal c-Myc epitope ...

Exploring the structure and function of Thermotoga ...

Apr 12, 2013·The crystal structure of TmCorA (Thermotoga maritima CorA) was the first divalent cation transporter structure that became available, presenting a funnel-shaped structure and an apparently closed hydrophobic channel [9–11].Later, the crystal structure of another Mg 2+ transporter from the MgtE family was reported [], which also presented a closed conformation.

An optimized protocol for protein purification in cultured ...

Jan 18, 2007·Kapust, R.B. et al. Tobacco etch virus protease: mechanism of autolysis and rational design of stable mutants with wild-type catalytic proficiency. Protein Eng. 14 , 993–1000 (2001).

urn:nbn:se:kth:diva-18191 : Exploring the activity of ...

Tobacco etch virus (TEV) protease is generally used to remove affinity tags from target proteins. It has been reported that some detergents inhibit the activity of this protease, and therefore should be avoided when removing affinity tags from membrane proteins. The aim of this study was to explore and evaluate this further.

Exploring the activity of tobacco etch virus protease in ...

Tobacco etch virus (TEV) protease is generally used to remove affinity tags from target proteins. It has been reported that some detergents inhibit the activity of this protease, and therefore ...

IJMS | Free Full-Text | Green Fluorescent Protein (GFP ...

The intrinsic kinase activity of AgrC in various detergent micelles was measured by ... H.; Eshaghi, S. Exploring the activity of tobacco etch virus protease in detergent solutions. Anal. Biochem 2008, 382, 69–71. [Google Scholar] Mohanty, A.K.; Simmons, C.R.; Wiener, M. Inhibition of tobacco etch virus protease activity by detergents. ...

IJMS | Free Full-Text | Green Fluorescent Protein (GFP ...

The intrinsic kinase activity of AgrC in various detergent micelles was measured by ... H.; Eshaghi, S. Exploring the activity of tobacco etch virus protease in detergent solutions. Anal. Biochem 2008, 382, 69–71. [Google Scholar] Mohanty, A.K.; Simmons, C.R.; Wiener, M. Inhibition of tobacco etch virus protease activity by detergents. ...

Frontiers | Easy Synthesis of Complex Biomolecular ...

There is a preference for working with an affinity tag at the C-terminus to make sure that only full-length proteins are purified when translation is incomplete. Tags can be eliminated after purification by insertion of an enzymatic cleavage site for thrombin or Tobacco Etch Virus (TEV) proteases (Waugh, 2011).

Quantitative Evaluation of Viral Protein Binding to ...

There is significant interest in developing analytical methods to characterize molecular recognition events between proteins and phosphoinositides, which are a medically important class of carbohydrate-functionalized lipids. Within this scope, one area of high priority involves quantitatively evaluating drug candidates that pharmacologically inhibit protein–phosphoinositide interactions.

Exploring the activity of tobacco etch virus protease in ...

Jan 30, 2017·Exploring the activity of tobacco etch virus protease in detergents Mezher, Amar () MOBK01 20162 Degree Projects in Molecular Biology. Mark; Popular Abstract Exploring the potential of biotechnological Tools

The high-throughput production of membrane proteins ...

Many proteases used to cleave MP fusion tags have reduced efficiency in the presence of some detergents [59, 60]. We favour the Tobacco etch virus (TEV) and Human rhinovirus (HRV) 3C proteases as both have stringent sequence specificity, are functional at 4°C and are easily producible at scale in the laboratory.

UNDERSTANDING STRUCTURE AND FUNCTION OF …

Exploring the activity of tobacco etch virus protease in detergent solutions. Anal Biochem. 382:69-71. II. Martinez Molina D¤, Lundbäck AK¤, Niegowski D and Eshaghi S. 2008. Expression and purification of the recombinant membrane protein YidC: A case study for increased stability and solubility. Protein Expr Purif. 62:49-52. III.

West Nile virus protease activity in detergent solutions ...

Jan 03, 2013·Exploring the activity of tobacco etch virus protease in detergent solutions. Lundback AK , van den Berg S , Hebert H , Berglund H , Eshaghi S Anal Biochem, (1):69-71 2008

Exploring the activity of tobacco etch virus protease in ...

Abstract. Tobacco etch virus (TEV) protease is generally used to remove affinity tags from target proteins. It has been reported that some detergents inhibit the activity of this protease, and therefore should be avoided when removing affinity tags from membrane proteins. The aim of this study was to explore and evaluate this further.

Engineering vanilloid-sensitivity into the rat TRPV2 ...

May 13, 2016·The protein was then eluted in 250 mM imidazole, and cleaved during overnight dialysis at 4°C with tobacco etch virus (TEV) protease (2 mg TEV/1 mg protein as estimated by fluorescence) in 3,500 MWCO dialysis tubing with stirring in 1.5 L dialysis buffer (150 mM NaCl, 20 mM Tris, 3x CMC detergent).

Structure-Function Analysis of the Bestrophin Family of ...

Mapping of transmembrane topography by insertion of N-linked glycosylation sites and tobacco etch virus protease cleavage sites provides evidence for cytosolic N and C termini and an unexpected transmembrane topography with at least three extracellular loops that include positions 60–63, 212–227, and 261–267. These experiments provide the ...