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A membrane protein based biosensor: Use of a phosphate – H+ symporter membrane protein (Pho84) in the sensing of phosphate ions.
Shabana, Basheer and Samyn, Dieter and Hedström, Martin and Thakur, M. S. (2011) A membrane protein based biosensor: Use of a phosphate – H+ symporter membrane protein (Pho84) in the sensing of phosphate ions. Biosensors and Bioelectronics, 27. pp. 58-63.
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Biosensors and Bioelectronics 2011 27(1) 58-.pdf - Published Version Restricted to Registered users only Download (767kB) |
Abstract
A label free biosensor for direct detection of inorganic phosphate based on potential-step capacitance measurements has been developed. The high-affinity Pho84 plasma membrane phosphate/proton symporter of Saccharomyces cerevisiae was used as a sensing element. Heterologously expressed and purified Pho84 protein was immobilized on a self-assembled monolayer (SAM) on a capacitance electrode. Changes in capacitance were recorded upon exposure to phosphate compared to the control substance, phosphate analogue methylphosphonate. Hence, even without the explicit use of lipid membranes, the Pho84 membrane protein could retain its capacity of selective substrate binding, with a phosphate detection limit in the range of the apparent in vivo Km. A linear increase in capacitance was monitored in the phosphate concentration range of 5–25 �M. The analytical response of the capacitive biosensor is in agreement with that the transporter undergoes significant conformational changes upon exposure to inorganic phosphate, while exposure to the analogue only causes minor responses.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | Capacitive biosensor, Membrane protein, Pho84 Phosphate, Symporter |
| Subjects: | 600 Technology > 05 Chemical engineering > 01 Biotechnology and Bioengineering |
| Divisions: | Fermentation Technology and Bioengineering |
| Depositing User: | Food Sci. & Technol. Information Services |
| Date Deposited: | 10 Feb 2012 04:22 |
| Last Modified: | 11 Jun 2019 05:57 |
| URI: | http://ir.cftri.res.in/id/eprint/10594 |
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