Analysis of an enzyme-substrate complex by X-ray crystallography and transferred nuclear Overhauser enhancement measurements: porcine pancreatic elastase and a hexapeptide.
The hexapeptide substrate Thr-Pro-nVal-NMeLeu-Tyr-Thr reacts with porcine pancreatic elastase sufficiently slowly that accelerated crystallographic data collection procedures and two-dimensional transferred nuclear Overhauser enhancement measurements could be used to study the geometry of binding. Both studies report a time-averaged population of the Michaelis complex state, prior to proteolysis. This result provides an important data point along the reaction coordinate pathway for serine proteases. Crystallographic data to 1.80-A resolution were used in the structure analysis with refinement to an R-factor of 0.19.
Meyer
E F
EF
Department of Biochemistry and Biophysics, Texas A&M University, College Station 77843-2128.
Clore
G M
GM
Gronenborn
A M
AM
Hansen
H A
HA
D000818:Animals; D001665:Binding Sites; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008958:Models, Molecular; D009842:Oligopeptides / Q000378:metabolism*; D010179:Pancreas / Q000201:enzymology*; D010196:Pancreatic Elastase / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D013552:Swine; D014961:X-Ray Diffraction / Q000379:methods
D009842:Oligopeptides; D010196:Pancreatic Elastase