Department of Chemistry and Biochemistry, University of California, Los Angeles, 607 Charles E. Young Drive East, Los Angeles, CA 90095 (USA); California NanoSystems Institute, University of California, Los Angeles, 570 Westwood Plaza, Los Angeles, CA 90095 (USA).
Chembiochem. 2013 Dec 16;14(18):2464-71. doi: 10.1002/cbic.201300478. Epub 2013 Oct 25.
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 proteins-horse heart myoglobin (HHMb) containing an N-terminal α-oxoamide and a red fluorescent protein (mCherry-CVIA) with a C-terminal alkyne-were immobilized by incubation onto respective stamped functionalized alkanethiol patterns. Pattern formation was confirmed by fluorescence microscopy.
本文采用功能化烷硫醇的微接触印刷和蛋白质的定点修饰相结合的方法，通过肟或铜催化的炔基-叠氮点击化学反应，将蛋白质选择性地固定在金表面上。合成了两种能够进行点击反应的分子，即氨氧基功能化的烷硫醇和叠氮功能化的烷硫醇，并通过红外光谱确认了金表面的自组装单分子层（SAM）的形成。然后，通过微接触印刷将烷硫醇分别图案化到金表面上。通过孵育到各自的印压功能化烷硫醇图案上，将含有 N 端α-氧代酰胺的定点修饰的蛋白质-马心血红蛋白（HHMb）和带有 C 端炔基的红色荧光蛋白（mCherry-CVIA）固定在金表面上。通过荧光显微镜确认了图案的形成。
