Visualizing enzyme and ribozyme intermediates using fast diffraction and reaction trapping methods

摘要

Using mutatagenesis, steady-state trapping, and photoactivation of caged substrates in two separate series of experiments, we have determined the structures of three sequential intermediates formed by the enzyme isocitrate dehydrogenase. Using pH triggering and physical trapping (flash-cooling), we have determined the structure of an unmodified, fully reactive hammerhead ribozyme construct at two separate stages of the autolytic cleavage reaction, and have proposed a structural mechanism for transition state formation. [References: 16]