ArticleLangmuir : the ACS journal of surfaces and colloids2016
Nonfouling NTA-PEG-Based TEM Grid Coatings for Selective Capture of Histidine-Tagged Protein Targets from Cell Lysates.
Article in Langmuir : the ACS journal of surfaces and colloids, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed.
- Open architecture of archaea MCM and dsDNA complexes resolved using monodispersed streptavidin affinity CryoEM.Nature communications · 2024Article
- An immobilized antibody-based affinity grid strategy for on-grid purification of target proteins enables high-resolution cryo-EM.Communications biology · 2024Article
- Polyelectrolyte coating of cryo-EM grids improves lateral distribution and prevents aggregation of macromolecules.Acta crystallographica. Section D, Structural biology · 2022Article
- Perspective: Biochemical and Physical Constraints Associated With Preparing Thin Specimens for Single-Particle Cryo-EM.Frontiers in molecular biosciences · 2022Article
- The His-tag as a decoy modulating preferred orientation in cryoEM.Frontiers in molecular biosciences · 2022Article
- Affinity Capture of p97 with Small-Molecule Ligand Bait Reveals a 3.6 Å Double-Hexamer Cryoelectron Microscopy Structure.ACS nano · 2021Article
- Nanodiscs: A toolkit for membrane protein science.Protein science : a publication of the Protein Society · 2021Review
- General and robust covalently linked graphene oxide affinity grids for high-resolution cryo-EM.Proceedings of the National Academy of Sciences of the United States of America · 2020Article
- Biological Applications at the Cutting Edge of Cryo-Electron Microscopy.Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada · 2018Review
- Cryo-EM in drug discovery: achievements, limitations and prospects.Nature reviews. Drug discovery · 2018Review
- Approaches to altering particle distributions in cryo-electron microscopy sample preparation.Acta crystallographica. Section D, Structural biology · 2018Review
- Monolayer-crystal streptavidin support films provide an internal standard of cryo-EM image quality.Journal of structural biology · 2017Article
- Atomic cryo-EM structures of viruses.Current opinion in structural biology · 2017Review
- Cryo-EM: beyond the microscope.Current opinion in structural biology · 2017Review
- The potential use of single-particle electron microscopy as a tool for structure-based inhibitor design.Acta crystallographica. Section D, Structural biology · 2017Article
- Opinion: hazards faced by macromolecules when confined to thin aqueous films.Biophysics reports · 2017Review
- Antibody-Based Affinity Cryoelectron Microscopy at 2.6-Å Resolution.Structure (London, England : 1993) · 2016Article
- Selective Capture of Histidine-tagged Proteins from Cell Lysates Using TEM grids Modified with NTA-Graphene Oxide.Scientific reports · 2016Article
- The changing landscape of membrane protein structural biology through developments in electron microscopy.Molecular membrane biology · 2016Review
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10 authors.
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Abstract
We report the preparation and performance of TEM grids bearing stabilized nonfouling lipid monolayer coatings. These films contain NTA capture ligands of controllable areal density at the distal end of a flexible poly(ethylene glycol) 2000 (PEG2000) spacer to avoid preferred orientation of surface-bound histidine-tagged (His-tag) protein targets. Langmuir-Schaefer deposition at 30 mN/m of mixed monolayers containing two novel synthetic lipids-1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[(5-amido-1-carboxypentyl)iminodiacetic acid]polyethylene glycolamide 2000) (NTA-PEG2000-DSPE) and 1,2-(tricosa-10',12'-diynoyl)-sn-glycero-3-phosphoethanolamine-N-(methoxypolyethylene glycolamide 350) (mPEG350-DTPE)-in 1:99 and 5:95 molar ratios prior to treatment with a 5 min, 254 nm light exposure was used for grid fabrication. These conditions were designed to limit nonspecific protein adsorption onto the stabilized lipid coating by favoring the formation of a mPEG350 brush layer below a flexible, mushroom conformation of NTA-PEG2000 at low surface density to enable specific immobilization and random orientation of the protein target on the EM grid. These grids were then used to capture His6-T7 bacteriophage and RplL from cell lysates, as well as purified His8-green fluorescent protein (GFP) and nanodisc solubilized maltose transporter, His6-MalFGK2. Our findings indicate that TEM grid supported, polymerized NTA lipid monolayers are capable of capturing His-tag protein targets in a manner that controls their areal densities, while efficiently blocking nonspecific adsorption and limiting film degradation, even upon prolonged detergent exposure.
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