Structural Enzymology

A long time ago, when the lab first started, we focused a great deal of effort into the development of methods for time-resolved crystallographic analyses of enzymatic turnover and reaction pathways.  This included fast Laue diffraction experiments, various ways to trigger and synchronize  turnover in the crystal, and the exploitation of a variety of methods for trapping intermediate species.  This area of technological development is no longer a strong focus of the lab, although we still make use of these methods when appropriate as part of our bread and butter projects on nucleic acid enzymes described in the other project links.

These days, our lab occasionally enters into short-term collaborations with local biochemists and enzymologists where specific ‘targets of opportunity’ that present interesting research problems for students and postdocs.  In many cases, these projects allow such trainees based in collaborator’s laboratories to spend some time learning basic methods of structural analyses.  While many of these experiments are clearly different from our primary interests , they offer the benefit of exposing our ‘regular’ students to detailed discussions of unique research activities.

Often, the methods that are introduced into our laboratory through these short-term experiments appear in modified form in our primary research activities.  For example, a short-term project on the structure and allosteric regulation of yeast pyruvate kinase introduced the laboratory to the use of high throughput robotic liquid handling systems for the identification of enzyme deregulators. This methodology and instrumentation is now being sometimes exploited by our laboratory both for drug screening experiments , and in a very different format for the selection of gene targeting endonuclease variants.

Laue diffraction
Laue diffraction on cover of Synchotron Radiation News, March/April 1992, Vol. 4, No. 2.

Future goals in this area

Who knows?  We like to go in new directions when the opportunity arises, and are always open to new experimental questions and approaches that we find interesting and educational.