Structural Aspects of Protein-Metal Recognition and Discrimination

摘要

This project utilized the zinc enzyme human carbonic anhydrase II (CAII) as a paradigm for dissecting and understanding the structural basis of protein-transition metal recognition and discrimination. We have successfully redesigned the zinc binding site of CAII so that protein-metal affinity can be modulated by design, by factors of 10 from micromolar to femtomolar affinity. We have explored the binding of zinc and copper to engineered CAIIs using high resolution X-ray crystallographic methods, and we have developed a zinc-sensing scheme using fluorescence anisotropy and a designed sulfonamide inhibitor.