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A membrane protein based biosensor: Use of a phosphate - H(+) symporter membrane protein (Pho84) in the sensing of phosphate ions.

Basheer, Shabana LU ; Samyn, Dieter; Hedström, Martin LU ; Thakur, Munna Singh; Persson, Bengt and Mattiasson, Bo LU (2011) In Biosensors & Bioelectronics 27(1). p.58-63
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 K(m).... (More)
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 K(m). A linear increase in capacitance was monitored in the phosphate concentration range of 5-25μM. 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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Membrane protein, Capacitive biosensor, Pho84, Phosphate, Symporter
in
Biosensors & Bioelectronics
volume
27
issue
1
pages
58 - 63
publisher
Elsevier
external identifiers
  • wos:000295953300009
  • scopus:79960923460
ISSN
1873-4235
DOI
10.1016/j.bios.2011.06.024
language
English
LU publication?
yes
id
bf2c2d65-a66a-428e-9dc0-5ff9a9ee016f (old id 2058286)
date added to LUP
2011-07-27 09:40:01
date last changed
2017-01-01 06:28:44
@article{bf2c2d65-a66a-428e-9dc0-5ff9a9ee016f,
  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 K(m). A linear increase in capacitance was monitored in the phosphate concentration range of 5-25μM. 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.},
  author       = {Basheer, Shabana and Samyn, Dieter and Hedström, Martin and Thakur, Munna Singh and Persson, Bengt and Mattiasson, Bo},
  issn         = {1873-4235},
  keyword      = {Membrane protein,Capacitive biosensor,Pho84,Phosphate,Symporter},
  language     = {eng},
  number       = {1},
  pages        = {58--63},
  publisher    = {Elsevier},
  series       = {Biosensors & Bioelectronics},
  title        = {A membrane protein based biosensor: Use of a phosphate - H(+) symporter membrane protein (Pho84) in the sensing of phosphate ions.},
  url          = {http://dx.doi.org/10.1016/j.bios.2011.06.024},
  volume       = {27},
  year         = {2011},
}