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Proceedings Paper

Fluorescence-based sensing of transition metal ions by a carbonic anhydrase transducer with a tethered fluorophore
Author(s): Richard B. Thompson; Badri P. Maliwal; Carol A. Fierke
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Paper Abstract

We have demonstrated that free metal ions such as Zn(II) can be determined by fuorescence anisotropy (polarization) using an apometalloenzyme, carbonic anhydrase II, and a fluorescent aryl sulfonamide inhibitor of the enzyme whose affinity for the enzyme is metal-dependent. We felt that attaching the fluorescent aryl sulfonamide to the protein would provide a similar response, while avoiding problems of disproportionation of the inhibitor and protein. In fact a tethered aryl sulfonamide ABD-T gave very good results: Zn(II) and Cu(II) at picomolar levels and Co(II), Cd(II), and Ni(II) at nanomolar levels can all be determined by changes in fluorescence intensity, anisotropy, and lifetime using visible excitation sources. Implications of these results are discussed.

Paper Details

Date Published: 3 May 1999
PDF: 8 pages
Proc. SPIE 3602, Advances in Fluorescence Sensing Technology IV, (3 May 1999); doi: 10.1117/12.347524
Show Author Affiliations
Richard B. Thompson, Univ. of Maryland School of Medicine (United States)
Badri P. Maliwal, Univ. of Maryland School of Medicine (United States)
Carol A. Fierke, Duke Univ. Medical Ctr. (United States)

Published in SPIE Proceedings Vol. 3602:
Advances in Fluorescence Sensing Technology IV
Joseph R. Lakowicz; Steven A. Soper; Richard B. Thompson, Editor(s)

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