We use and develop rapid-reaction kinetic equipment to measure protein-ligand interactions. See Molecular Enzymology Group . We mainly use fluorescence detection because of its excellent sensitivity. Probes we exploit include tryptophan, pyrene, GFP and a variety of fluorescent nucleotide analogues. We have developed a detailed reaction mechanism for the Dictyostelium myosin II  ATPase

This stopped-flow record shows a transient rapid decrease in tryptophan fluorescence on mixing Dictyostelium myosin II motor domain with ATP due to M⁺ATP + M*ATP formation (M⁺ATP more significant at lower temperatures), followed by a rise in fluorescence as the M*ATP <--> M.*ADP.Pi states requilibrate on hydrolysis of ATP. Relaxation methods (temperature and pressure jump) were required to resolve the M⁺ATP <--> M*ATP interconversion (which corresponds to switch 2 closing).

1lvk  X-RAY CRYSTAL STRUCTURE OF THE Mg 2'(3')-O-(N-METHYLANTHRANILOYL) NUCLEOTIDE BOUND TO DICTYOSTELIUM DISCOIDEUM MYOSIN MOTOR DOMAIN

1qkr  CRYSTAL STRUCTURE OF THE VINCULIN TAIL AND A PATHWAY FOR ACTIVATION

2k00  Solution structure of the talin F3 in complex with layilin cytodomain

1c1v  The 1.5 A Crystal structure of Ca²⁺-bound S100A4

3MYH 3MYK 3MYL  Ser236Ala mutant of Dictyostelium myosin II motor domain