Catalytic promiscuity in a protein superfamily

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Understanding of enzyme catalysis remains a daunting challenge, despite intense research efforts in basic and applied research. Our understanding certainly fails the most severe test - that of making catalysts that meet the efficiency of natural enzymes. This proposal addresses the determinants of catalytic efficiency in hydrolytic enzymes, using as a test bed a superfamily of catalysts that are connected by structural homology as members of the alkaline phosphatase superfamily as well as often reciprocal and crosswise catalytic promiscuity. Starting from our recent identification of one remarkable example for multiple promiscuity and high efficiency (Proc. Natl. Acad. Sci - in press, doi/10.1073/pnas.0903951107), we propose to address the factors that determine selectivity and efficiency, by structural and kinetic analysis and by directed evolution.