Chemoselective immobilization of proteins by microcontact printing and bio-orthogonal click reactions
| Autor: | Dayanara P. Parra, Erhan Bat, Jonathan K. Dozier, Mark D. Distefano, Zachary P. Tolstyka, Bruce Dunn, Wade Richardson, Caitlin J. Stevens, Heather D. Maynard |
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| Rok vydání: | 2013 |
| Předmět: |
Azides
Surface Properties Biochemistry Catalysis Article Monolayer Polymer chemistry Alkanes Oximes Fluorescence microscope Molecule Animals Horses Sulfhydryl Compounds Chemoselectivity Molecular Biology Luminescent Proteins Chemistry Myoglobin Organic Chemistry Immobilized Proteins Microcontact printing Alkynes Click chemistry Molecular Medicine Click Chemistry Self-assembly Gold Copper |
| Zdroj: | Chembiochem : a European journal of chemical biology. 14(18) |
| ISSN: | 1439-7633 |
| Popis: | Herein, a combination of microcontact printing of functionalized alkanethiols and site-specific modification of proteins is utilized to chemoselectively immobilize proteins onto gold surfaces either by oxime or copper catalyzed alkyne-azide click chemistry. Two molecules capable of click reactions, an aminooxy-functionalized alkanethiol and an azide-functionalized alkanethiol, were synthesized, and self-assembled monolayer (SAM) formation on gold was confirmed by IR spectroscopy. The alkanethiols were then individually patterned onto gold surfaces by microcontact printing. Site-specifically modified proteins, horse heart myoglobin (HHMb) containing an N-terminal α-oxoamide and a red-fluorescent protein (mCherry-CVIA) with a C-terminal alkyne, respectively were immobilized by incubation onto the stamped functionalized alkanethiol patterns. Pattern formation was confirmed by fluorescence microscopy. |
| Databáze: | OpenAIRE |
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