Effect of Temperature on Stability and Activity of Elongation Factor 2 Proteins from Antarctic and Thermophilic Methanogens

• Published in: Journal of Bacteriology
• Main Authors: Thomas, Torsten, Cavicchioli, Ricardo
• Format: Article in Journal/Newspaper
• Language: English
• Published: American Society for Microbiology 2000
• Subjects: Antarctic; The Antarctic; Antarc*
• Online Access: http://dx.doi.org/10.1128/jb.182.5.1328-1332.2000; https://journals.asm.org/doi/pdf/10.1128/JB.182.5.1328-1332.2000

ABSTRACT Despite the presence and abundance of archaea in low-temperature environments, little information is available regarding their physiological and biochemical properties. In order to investigate the adaptation of archaeal proteins to low temperatures, we purified and characterized the elongation factor 2 (EF-2) protein from the Antarctic methanogen Methanococcoides burtonii , which was expressed in Escherichia coli , and compared it to the recombinant EF-2 protein from a phylogenetically related thermophile, Methanosarcina thermophila . Using differential scanning calorimetry to assess protein stability and enzyme assays for the intrinsic GTPase activity, we identified biochemical and biophysical properties that are characteristic of the cold-adapted protein. This includes a higher activity at low temperatures caused by a decrease of the activation energy necessary for GTP hydrolysis and a decreased activation energy for the irreversible denaturation of the protein, which indicates a less thermostable structure. Comparison of the in vitro properties of the proteins with the temperature-dependent characteristics of growth of the organisms indicates that additional cytoplasmic factors are likely to be important for the complete thermal adaptation of the proteins in vivo. This is the first study to address thermal adaptation of proteins from a free-living, cold-adapted archaeon, and our results indicate that the ability of the Antarctic methanogen to adapt to the cold is likely to involve protein structural changes.