Controlling protein surface orientation by strategic placement of oligo-histidine tags
| Autor: | Christian Blum, Pierre Andre Cazade, Shane O'Mahony, Pascal Jonkheijm, Jord C. Prangsma, Jurriaan Huskens, Jordi Cabanas-Danés, Damien Thompson, Vinod Subramaniam, Eldrich E. Tromp, Dorothee Wasserberg |
|---|---|
| Přispěvatelé: | German Academic Exchange Service, Stichting Technische Wetenschappen, ERC, SFI, Molecular Nanofabrication, Nanobiophysics, Executive board Vrije Universiteit |
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
Nitrilotriacetic Acid
Surface Properties General Physics and Astronomy Infrared spectroscopy 02 engineering and technology Molecular Dynamics Simulation 010402 general chemistry 01 natural sciences Article chemistry.chemical_compound Nickel Fluorescence microscope Animals General Materials Science Histidine Surface plasmon resonance protein immobilization Monolayers Chemistry Molecular dynamics simulations General Engineering Nitrilotriacetic acid Self-assembly molecular dynamics simulations self-assembly multivalency 021001 nanoscience & nanotechnology Fluorescence 0104 chemical sciences Crystallography Luminescent Proteins Immobilized Proteins Sea Anemones Protein immobilization monolayers Multivalency 0210 nano-technology Oligopeptides Fluorescence anisotropy |
| Zdroj: | ACS nano, 11(9), 9068-9083. American Chemical Society ACS Nano, 11(9), 9068-9083. American Chemical Society ACS Nano Wasserberg, D, Cabanas-Danés, J, Prangsma, J, O'Mahony, S, Cazade, P A, Tromp, E, Blum, C, Thompson, D, Huskens, J, Subramaniam, V & Jonkheijm, P 2017, ' Controlling Protein Surface Orientation by Strategic Placement of Oligo-Histidine Tags ', ACS Nano, vol. 11, no. 9, pp. 9068-9083 . https://doi.org/10.1021/acsnano.7b03717 |
| ISSN: | 1936-0851 |
| DOI: | 10.1021/acsnano.7b03717 |
| Popis: | We report oriented immobilization of proteins using the standard hexahistidine (His6)-Ni2+:NTA (nitrilotriacetic acid) methodology, which we systematically tuned to give control of surface coverage. Fluorescence microscopy and surface plasmon resonance measurements of self-assembled monolayers (SAMs) of red fluorescent proteins (TagRFP) showed that binding strength increased by 1 order of magnitude for each additional His6-tag on the TagRFP proteins. All TagRFP variants with His6-tags located on only one side of the barrel-shaped protein yielded a 1.5 times higher surface coverage compared to variants with His6-tags on opposite sides of the so-called β-barrel. Time-resolved fluorescence anisotropy measurements supported by polarized infrared spectroscopy verified that the orientation (and thus coverage and functionality) of proteins on surfaces can be controlled by strategic placement of a His6-tag on the protein. Molecular dynamics simulations show how the differently tagged proteins reside at the surface in "end-on" and "side-on" orientations with each His6-tag contributing to binding. Also, not every dihistidine subunit in a given His6-tag forms a full coordination bond with the Ni2+:NTA SAMs, which varied with the position of the His6-tag on the protein. At equal valency but different tag positions on the protein, differences in binding were caused by probing for Ni2+:NTA moieties and by additional electrostatic interactions between different fractions of the β-barrel structure and charged NTA moieties. Potential of mean force calculations indicate there is no specific single-protein interaction mode that provides a clear preferential surface orientation, suggesting that the experimentally measured preference for the end-on orientation is a supra-protein, not a single-protein, effect. |
| Databáze: | OpenAIRE |
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