Specific adsorption of histidine-tagged proteins on silica surfaces modified with Ni2+/NTA-derivatized poly(ethylene glycol)
Silica surfaces modified with nitrilotriacetic acid (NTA)-polyethylene glycol (PEG) derivatives were used to immobilize hexahistidine-tagged green fluorescent protein (His6-GFP), biotin/streptavidin-AlexaFluor555 (His6-biotin/SA-AF), and gramicidin A-containing vesicles (His6-gA). Three types of sur...
| Veröffentlicht in: | Langmuir : the ACS journal of surfaces and colloids. - 1992. - 23(2007), 11 vom: 22. Mai, Seite 6281-8 |
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| 1. Verfasser: | |
| Weitere Verfasser: | , , , , , , , |
| Format: | Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
2007
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| Zugriff auf das übergeordnete Werk: | Langmuir : the ACS journal of surfaces and colloids |
| Schlagworte: | Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S. Lipid Bilayers Proteins Polyethylene Glycols 3WJQ0SDW1A Histidine 4QD397987E Silicon Dioxide mehr... |
| Zusammenfassung: | Silica surfaces modified with nitrilotriacetic acid (NTA)-polyethylene glycol (PEG) derivatives were used to immobilize hexahistidine-tagged green fluorescent protein (His6-GFP), biotin/streptavidin-AlexaFluor555 (His6-biotin/SA-AF), and gramicidin A-containing vesicles (His6-gA). Three types of surface-reactive PEG derivatives-NTA-PEG3400-Si(OMe)3, NTA-PEG3400-vinylsulfone, and mPEG5000-Si(OMe)3 (control)-were grafted onto silica and tested for their ability to capture His6-tag species via His6/Ni2+/NTA chelation. The composition and thicknesses of the PEG-modified surfaces were characterized using X-ray photoelectron spectroscopy, contact angle, and ellipsometry. Protein capture efficiencies of the NTA-PEG-grafted surfaces were evaluated by measuring fluorescence intensities of these surfaces after exposure to His6-tag species. XPS and ellipsometry data indicate that surface adsorption occurs via specific interactions between the His6-tag and the Ni2+/NTA-PEG-grafted surface. Protein immobilization was most effective for NTA-PEG3400-Si(OMe)3-modified surfaces, with maximal areal densities achieved at 45 pmol/cm2 for His6-GFP and 95 fmol/cm2 for His6-biotin/SA-AF. Lipid vesicles containing His6-gA in a 1:375 gA/lipid ratio could also be immobilized on Ni2+/NTA-PEG3400-Si(OMe)3-modified surfaces at 0.5 mM total lipid. Our results suggest that NTA-PEG-Si(OMe)3 conjugates may be useful tools for immobilizing His6-tag proteins on solid surfaces to produce protein-functionalized surfaces |
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| Beschreibung: | Date Completed 11.07.2007 Date Revised 08.01.2019 published: Print-Electronic Citation Status MEDLINE |
| ISSN: | 1520-5827 |