Gastric ATPase phosphorylation/dephosphorylation monitored by new FTIR-based BIA–ATR biosensors. Issue 3 (2010)

• Record Type: Journal Article
• Authors: Goldsztein Goldsztein, A. A.; Babar Babar, S. S.; Voué Voué, M. M.; De Coninck De Coninck, J. J.; Conti Conti, J. J.; Marchand-Brynaert Marchand-Brynaert, J. J.; Devouge Devouge, S. S.; Homblé Homblé, F. F.; Goormaghtigh Goormaghtigh, E. E.
• Is Part Of: Spectroscopy. Volume 24:Issue 3/4(2010)
• Page Start: 257
• Page End: 260
• Publication Date: 2010
• Subjects: Gastric ATPase -- BIA–ATR biosensors -- FTIR -- phosphorylation
• DOI: 10.3233/SPE-2010-0402
• Languages: English
• ISSNs: 0712-4813

Abstract

Abstract : 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.