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Zinc Biochemistry, Physiology, and Homeostasis pp 19–36Cite as

Fluorescence-based biosensing of zinc using carbonic anhydrase

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Abstract

Measurement of free zinc levels and imaging of zinc fluxes remains technically difficult due to low levels and the presence of interfering cations such as Mg and Ca. We have developed a series of fluorescent zinc indicators based on the superb sensitivity and selectivity of a protein, human apo-carbonic anhydrase II, for Zn(II). These indicators transduce the level of free zinc as changes in intensity, wavelength ratio, lifetime, and/or anisotropy; the latter three approaches permit quantitative imaging of zinc levels in the microscope. A unique attribute of sensors incorporating biological macromolecules as transducers is their capability for modification by site-directed mutagenesis. Thus we have produced variants of carbonic anhydrase with improved affinity for zinc, altered selectivity, and enhanced binding kinetics, all of which are difficult to modify in small molecule indicators.

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Author information

Authors and Affiliations

  1. Departments of Chemistry and Biochemistry, University of Michigan, Ann Arbor, Michigan, USA

    Carol A. Fierke

  2. Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland, USA

    Richard B. Thompson

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  1. Carol A. Fierke
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  2. Richard B. Thompson
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Editors and Affiliations

  1. Center for Biochemical and Biophysical Sciences and Medicine, Harvard Medical School, Boston, MA, USA

    W. Maret

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Fierke, C.A., Thompson, R.B. (2001). Fluorescence-based biosensing of zinc using carbonic anhydrase. In: Maret, W. (eds) Zinc Biochemistry, Physiology, and Homeostasis. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-3728-9_2

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  • DOI: https://doi.org/10.1007/978-94-017-3728-9_2

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  • Print ISBN: 978-90-481-5916-1

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