Fluorescence-based sensing of transition metal ions by a carbonic anhydrase transducer with a tethered fluorophore

Richard B. Thompson; Badri P. Maliwal; Carol A. Fierke

doi:10.1117/12.347524

3 May 1999 Fluorescence-based sensing of transition metal ions by a carbonic anhydrase transducer with a tethered fluorophore

Richard B. Thompson, Badri P. Maliwal, Carol A. Fierke

Author Affiliations +

Proceedings Volume 3602, Advances in Fluorescence Sensing Technology IV; (1999) https://doi.org/10.1117/12.347524
Event: BiOS '99 International Biomedical Optics Symposium, 1999, San Jose, CA, United States

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.

Citation Download Citation

Richard B. Thompson, Badri P. Maliwal, and Carol A. Fierke "Fluorescence-based sensing of transition metal ions by a carbonic anhydrase transducer with a tethered fluorophore", Proc. SPIE 3602, Advances in Fluorescence Sensing Technology IV, (3 May 1999); https://doi.org/10.1117/12.347524

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