Adaptation to high temperatures through macromolecular dynamics by neutron scattering.
Work on the relationship between hyperthermophile protein dynamics, stability and activity is reviewed. Neutron spectroscopy has been applied to measure and compare the macromolecular dynamics of various hyperthermophilic and mesophilic proteins, under different conditions. First, molecular dynamics have been analyzed for the hyperthermophile malate dehydrogenase from Methanococcus jannaschii and a mesophilic homologue, the lactate dehydrogenase from Oryctolagus cunniculus (rabbit) muscle. The neutron scattering approach has provided independent measurements of the global flexibility and structural resilience of each protein, and it has been demonstrated that macromolecular dynamics represents one of the molecular mechanisms of thermoadaptation. The resilience was found to be higher for the hyperthermophilic protein, thus ensuring similar flexibilities in both enzymes at their optimal activity temperature. Second, the neutron method has been developed to quantify the average macromolecular flexibility and resilience within the natural crowded environment of the cell, and mean macromolecular motions have been measured in vivo in psychrophile, mesophile, thermophile and hyperthermophile bacteria. The macromolecular resilience in bacteria was found to increase with adaptation to high temperatures, whereas flexibility was maintained within narrow limits, independent of physiological temperature for all cells in their active state. Third, macromolecular motions have been measured in free and immobilized dihydrofolate reductase from Escherichia coli. The immobilized mesophilic enzyme has increased stability and decreased activity, so that its properties are changed to resemble those of a thermophilic enzyme. Quasi-elastic neutron scattering measurements have also been performed to probe the protein motions. Compared to the free enzyme, the average height of the activation free energy barrier to local motions was found to be increased by 0.54 kcal.mol(-1) in the immobilized dihydrofolate reductase, a value that is of the same order as expected from the theoretical rate equation.
Institut Laue-Langevin, Grenoble, France. v-tehei@ill.fr
Moeava
M
Tehei
Giuseppe
G
Zaccai
D000222:Adaptation, Physiological; D000818:Animals; D019843:Archaeal Proteins / Q000737:chemistry*; D006358:Hot Temperature; D007770:L-Lactate Dehydrogenase / Q000737:chemistry*; D017017:Methanococcus / Q000201:enzymology / Q000254:growth & development; D009124:Muscle Proteins / Q000737:chemistry; D033363:Neutron Diffraction / Q000379:methods*; D017510:Protein Folding; D011817:Rabbits
D019843:Archaeal Proteins; D009124:Muscle Proteins; D007770:L-Lactate Dehydrogenase