期刊
CHEMBIOCHEM
卷 15, 期 17, 页码 2529-2534出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.201402478
关键词
ATR-FTIR; biosensors; Channelrhodopsin 2; germanium; protein immobilization
资金
- Deutsche Forschungsgemeinschaft [SFB 642]
Protein immobilization on solid surfaces has become a powerful tool for the investigation of protein function. Physiologically relevant molecular reaction mechanisms and interactions of proteins can be revealed with excellent signal-to-noise ratio by vibrational spectroscopy (ATR-FTIR) on germanium crystals. Protein immobilization by thiol chemistry is well-established on gold surfaces, for example, for surface plasmon resonance. Here, we combine features of both approaches: a germanium surface functionalized with different thiols to allow specific immobilization of various histidine-tagged proteins with over 99% specific binding. In addition to FTIR, the surfaces were characterized by XPS and fluorescence microscopy. Secondary-structure analysis and stimulus-induced difference spectroscopy confirmed protein activity at the atomic level, for example, physiological cation channel formation of Channelrhodopsin 2.
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