Abstract
Introduction
Material & Methods
Results
Conclusion
Literature Cited
Acknowledgements

 NSF Student Research
Rhonda Moeller and Diana Markoch: Generation of Apo-Carbonic Anhydrase to Study the Mechanism of Zinc Transfer from Metallothionein to Apo-zinc Metalloproteins
Conclusions

The apo-form of Carbonic anhydrase can be successfully prepared by dialyzing the enzyme against dipicolinic acid for 3 hours. This procedure resulted in a 95.7% reduction of enzyme activity. Furthermore, reconstitution of the enzyme was possible by the addition of 0.2 μM of Zinc to the apoenzyme. Our results showed 44% reactivation of CA relative to the holoenzyme. Using this assay, 3.13 μg of Carbonic Anhydrase can be detected. Finally, the Vmax and Km values were calculated from Lineweaver-Burke plots for carbonic anhydrase and the substrate p-nitrophenyl acetate. From the linear regression, the Vmax was calculated to be 1.33mM of p-NPA/mg CA protein/ minute and the Km was 0.093mM p-NPA.