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Proceedings Paper

Optical sensing systems based on biomolecular recognition of recombinant proteins
Author(s): Lyndon L.E. Salins; Vesna Schauer-Vukasinovic; Sylvia Daunert
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Paper Abstract

SIte-directed mutagenesis and the associated site-specific fluorescent labeling of proteins can be used to rationally design reagentless fluorescent molecular senors. The phosphate binding protein (PBP) and calmodulin (CaM) bind to phosphate and calcium in a highly specific manner. These ions induce a hinge motion in the proteins, and the resultant conformational change constitutes the basis of the sensor development. By labeling each protein at a specific site with environment-sensitive fluorescent probes, these conformational changes can be monitored and related to the amount of analyte ion present. In this study, the polymerase chain reaction was used to construct mutants of PBP and CaM that have a single cysteine at positions 197 and 109, respectively. Each protein was site-specifically labeled through the sulfhydryl group of the introduced cysteine residue at a single location with an environment-sensitive fluorescent probe. Characterization of the steady-state fluorescence indicated an enhancement of signal intensity upon binding of the analyte ion. Highly sensitive and selective and selective sensing systems for phosphate and calcium were obtained by using this approach.

Paper Details

Date Published: 15 May 1998
PDF: 9 pages
Proc. SPIE 3270, Methods for Ultrasensitive Detection, (15 May 1998); doi: 10.1117/12.308378
Show Author Affiliations
Lyndon L.E. Salins, Univ. of Kentucky (United States)
Vesna Schauer-Vukasinovic, Univ. of Kentucky (United States)
Sylvia Daunert, Univ. of Kentucky (United States)

Published in SPIE Proceedings Vol. 3270:
Methods for Ultrasensitive Detection
Bryan L. Fearey, Editor(s)

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