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  Home > JCE Print > Journal of Chemical Education > Issues > 2004  > July  >
In the Laboratory
Removal of Zinc from Carbonic Anhydrase. A Kinetics Experiment for Upper-Level Chemistry Laboratories
Kathryn R. Williams and Bhavin Adhyaru
Department of Chemistry, University of Florida, Gainesville, FL 32611-7200
Cover
July 2004
Vol. 81 No. 7
p. 1045
Abstract
The Zn(II) ion in the active site of carbonic anhydrase can be removed by complexation with 2,6-pyridinedicarboxylate (dipicolinate). The kinetics of Zn(II) removal can be followed by two methods. In the standard experiment (suitable for students in biophysical chemistry, biochemistry, or instrumental analysis), aliquots of the reaction mixture are withdrawn at suitable times and tested for the residual enzymatic activity (p-nitrophenyl acetate hydrolysis). In an optional experiment (special project or for students in radiochemistry), the carbonic anhydrase is labeled with 65Zn. Aliquots of the reaction mixture are withdrawn and filtered to separate the liberated 65Zn, which is detected using either a liquid scintillation counter or a gamma counter.
Supplement
Instructions for students, including a prelab assignment, and notes for the instructor are available.
*  Contents JCE2004p1045W.doc (Microsoft Word)
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*  Citation
 Williams, Kathryn R.; Adhyaru, Bhavin. J. Chem. Educ. 2004 81 1045.
*  Keywords
 Biochemistry; Biophysical Chemistry; Enzymes; Kinetics; Laboratory Instruction; Nuclear / Radiochemistry; Zinc
*  History
 Created:
Last Updated:		 May 27, 2004
January 19, 2005
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