Carbonic anhydrase II-based metal ion sensing: Advances and new perspectives |
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| Authors: | Tamiika K Hurst Da Wang Richard B Thompson Carol A Fierke |
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| Institution: | 1. Department of Chemistry, The University of Michigan, 930 N University Ave, Ann Arbor, MI 48109, USA;2. Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, 108 N Greene St., Baltimore, Maryland 21201, USA;3. Department of Biological Chemistry, The University of Michigan, 930 N University Ave, Ann Arbor, MI 48109, USA |
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| Abstract: | Carbonic anhydrases are archetypical zinc metalloenzymes and as such, they have been developed as the recognition element of a family of fluorescent indicators (sensors) to detect metal ions, particularly Zn2+ and Cu2+. Subtle modification of the structure of human carbonic anhydrase II isozyme (CAII) alters the selectivity, sensitivity, and response time for these sensors. Sensors using CAII variants coupled with zinc-dependent fluorescent ligands demonstrate picomolar sensitivity, unmatched selectivity, ratiometric fluorescence signal, and near diffusion-controlled response times. Recently, these sensors have been applied to measuring the readily exchangeable concentrations of zinc in the cytosol and nucleus of mammalian tissue culture cells and concentrations of free Cu2+ in seawater. |
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| Keywords: | AF594 AlexaFluor 594 CAII carbonic anhydrase isozyme II DNSA dansylamide FRET Fö rster resonance energy transfer ICP-MS inductively coupled plasma mass spectrometry NTA nitrilotriacetic acid TSQ N-6-methoxy-8-quinolyl)-p-toluenesulfonamide XRF X-ray fluorescence spectrometry |
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