Gastric ATPase phosphorylation/dephosphorylation monitored by new FTIR-based BIA–ATR biosensors
| Autor: | Goldsztein, A., Babar, S., Voué, M., De Coninck, J., Conti, J., Marchand-Brynaert, J., Devouge, S., Homblé, F., Goormaghtigh, E. |
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| Rok vydání: | 2010 |
| Předmět: | |
| Zdroj: | Spectroscopy. 24:257-260 |
| ISSN: | 1875-922X 0712-4813 |
| Popis: | Biosensors are composite devices suitable for the investigation of receptor–ligand interactions. In this paper we present the specific application to a membrane embedded protein of a new sensor device, so-called BIA–ATR, based on Attenuated Total Reflection–Fourier Transform Infrared (ATR–FTIR) spectroscopy. It consists in a functionalised ATR germanium crystal whose surface has been covalently modified to adsorb a biomembrane. Detection of the ligand–receptor interaction is achieved using FTIR spectroscopy. We report the specific detection of the phosphorylation/dephosphorylation of the H+/K+gastric ATPase. The H+, K+-ATPase is a particularly large protein entity. This glycosylated protein contains more than 1300 residues and is embedded in a lipid membrane. Yet we demonstrate that the BIA–ATR sensor is capable of monitoring the binding of a single phosphate on such a large protein entity. Furthermore, we also demonstrate the potential of the approach to monitor the kinetics of binding and dissociation of the ligand. |
| Databáze: | OpenAIRE |
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