The molecular basis of the effect of temperature on enzyme activity.
Experimental data show that the effect of temperature on enzymes cannot be adequately explained in terms of a two-state model based on increases in activity and denaturation. The Equilibrium Model provides a quantitative explanation of enzyme thermal behaviour under reaction conditions by introducing an inactive (but not denatured) intermediate in rapid equilibrium with the active form. The temperature midpoint (Teq) of the rapid equilibration between the two forms is related to the growth temperature of the organism, and the enthalpy of the equilibrium (DeltaHeq) to its ability to function over various temperature ranges. In the present study, we show that the difference between the active and inactive forms is at the enzyme active site. The results reveal an apparently universal mechanism, independent of enzyme reaction or structure, based at or near the active site, by which enzymes lose activity as temperature rises, as opposed to denaturation which is global. Results show that activity losses below Teq may lead to significant errors in the determination of DeltaG*cat made on the basis of the two-state ('Classical') model, and the measured kcat will then not be a true indication of an enzyme's catalytic power. Overall, the results provide a molecular rationale for observations that the active site tends to be more flexible than the enzyme as a whole, and that activity losses precede denaturation, and provide a general explanation in molecular terms for the effect of temperature on enzyme activity.
Department of Biological Sciences, University of Waikato, Private Bag 3105, Hamilton, New Zealand.
Roy M
RM
Daniel
Michelle E
ME
Peterson
Michael J
MJ
Danson
Nicholas C
NC
Price
Sharon M
SM
Kelly
Colin R
CR
Monk
Cristina S
CS
Weinberg
Matthew L
ML
Oudshoorn
Charles K
CK
Lee
D020134:Catalytic Domain; D004798:Enzymes / Q000378:metabolism*; D007700:Kinetics; D008956:Models, Chemical; D011489:Protein Denaturation; D013696:Temperature; D013816:Thermodynamics; D044366:Transition Temperature
D004798:Enzymes