Dynamics of immobilized and native Escherichia coli dihydrofolate reductase by quasielastic neutron scattering.
The internal dynamics of native and immobilized Escherichia coli dihydrofolate reductase (DHFR) have been examined using incoherent quasielastic neutron scattering. These results reveal no difference between the high frequency vibration mean-square displacement of the native and the immobilized E. coli DHFR. However, length-scale-dependent, picosecond dynamical changes are found. On longer length scales, the dynamics are comparable for both DHFR samples. On shorter length scales, the dynamics is dominated by local jump motions over potential barriers. The residence time for the protons to stay in a potential well is tau = 7.95 +/- 1.02 ps for the native DHFR and tau = 20.36 +/- 1.80 ps for the immobilized DHFR. The average height of the potential barrier to the local motions is increased in the immobilized DHFR, and may increase the activation energy for the activity reaction, decreasing the rate as observed experimentally. These results suggest that the local motions on the picosecond timescale may act as a lubricant for those associated with DHFR activity occurring on a slower millisecond timescale. Experiments indicate a significantly slower catalytic reaction rate for the immobilized E. coli DHFR. However, the immobilization of the DHFR is on the exterior of the enzyme and essentially distal to the active site, thus this phenomenon has broad implications for the action of drugs distal to the active site.
Tehei
M
M
Department of Biological Sciences, University of Waikato, Hamilton, New Zealand. v-tehei@ill.fr
Smith
J C
JC
Monk
C
C
Ollivier
J
J
Oettl
M
M
Kurkal
V
V
Finney
J L
JL
Daniel
R M
RM
D001665:Binding Sites; D001703:Biophysics / Q000379:methods*; D002384:Catalysis; D004800:Enzymes, Immobilized; D004926:Escherichia coli / Q000737:chemistry* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry; D007700:Kinetics; D008956:Models, Chemical; D008958:Models, Molecular; D015233:Models, Statistical; D009502:Neutrons; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012542:Scattering, Radiation; D012822:Silicon Dioxide; D013762:Tetrahydrofolate Dehydrogenase / Q000737:chemistry*; D013997:Time Factors
D004800:Enzymes, Immobilized; D029968:Escherichia coli Proteins; D012822:Silicon Dioxide; D013762:Tetrahydrofolate Dehydrogenase