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Chemoselective Immobilization of Proteins by Microcontact Printing and Bio-orthogonal Click Reactions

Tolstyka, Zachary P.
Richardson, Wade
Bat, Erhan
Stevens, Caitlin J.
Parra, Dayanara P.
Dozier, Jonathan K.
Distefano, Mark D.
Dunn, Bruce
Maynard, Heather D.
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 were synthesized, an aminooxy-functionalized alkanethiol and an azide-functionalized alkanethiol, 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 proteinshorse heart myoglobin (HHMb) containing an N-terminal -oxoamide and a red fluorescent protein (mCherry-CVIA) with a C-terminal alkynewere immobilized by incubation onto respective stamped functionalized alkanethiol patterns. Pattern formation was confirmed by fluorescence microscopy.