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

Basheer, ShabanaLund University,Lunds universitet,Bioteknik,Centrum för tillämpade biovetenskaper,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Biotechnology,Center for Applied Life Sciences,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH (author)

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 5-25 mu 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.

Samyn, Dieter R.Linnéuniversitetet,Institutionen för naturvetenskap, NV,School of Natural Sciences, Linnaeus University, Kalmar, Sweden(Swepub:oru)drsn (author)
Hedström, MartinLund University,Lunds universitet,Bioteknik,Centrum för tillämpade biovetenskaper,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Biotechnology,Center for Applied Life Sciences,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH(Swepub:lu)biot-mhe (author)
Thakur, Munna SinghDepartment of Fermentation, Technology and Bioengineering, Central Food Technology Research Institute, Mysore, India (author)
Persson, Bengt L.,1951-Linnéuniversitetet,Institutionen för naturvetenskap, NV,Bengt Persson,School of Natural Sciences, Linnaeus University, Kalmar, Sweden; Lab. of Molecular Cell Biology, Institute of Botany and Microbiology, Katholieke Universiteit, Leuven, Belgium; Department of Molecular Microbiology, VIB, Leuven-Heverlee, Flanders, Belgium(Swepub:lnu)hpebe (author)
Mattiasson, BoLund University,Lunds universitet,Bioteknik,Centrum för tillämpade biovetenskaper,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Biotechnology,Center for Applied Life Sciences,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH(Swepub:lu)biot-bma (author)
BioteknikCentrum för tillämpade biovetenskaper (creator_code:org_t)

By the author/editor: Basheer, Shabana; Samyn, Dieter R.; Hedström, Martin; Thakur, Munna Si ...; Persson, Bengt L ...; Mattiasson, Bo

About the subject

NATURAL SCIENCES: NATURAL SCIENCES; and Biological Scien ...; and Biochemistry and ...

NATURAL SCIENCES: NATURAL SCIENCES; and Chemical Science ...; and Analytical Chemi ...

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