title: A thermodynamic comparison of mesophilic and thermophilic ribonucleases H.
abstract: The mechanisms by which thermophilic proteins attain their increased thermostability remain unclear, as usually the sequence and structure of these proteins are very similar to those of their mesophilic homologues. To gain insight into the basis of thermostability, we have determined protein stability curves describing the temperature dependence of the free energy of unfolding for two ribonucleases H, one from the mesophile Escherichia coli and one from the thermophile Thermus thermophilus. The circular dichroism signal was monitored as a function of temperature and guanidinium chloride concentration, and the resulting free energies of unfolding were fit to the Gibbs-Helmholtz equation to obtain a set of thermodynamic parameters for these proteins. Although the maximal stabilities for these proteins occur at similar temperatures, the heat capacity of unfolding for T. thermophilus RNase H is lower, resulting in a smaller temperature dependence of the free energy of unfolding and therefore a higher thermal melting temperature. In addition, the stabilities of these proteins are similar at the optimal growth temperatures for their respective organisms, suggesting that a balance of thermodynamic stability and flexibility is important for function.
authors: Department of Molecular and Cell Biology, University of California, Berkeley, California 94720-3206, USA.
authors: J
authors: J
authors: Hollien
authors: S
authors: S
authors: Marqusee
mesh_terms: D000595:Amino Acid Sequence; D002942:Circular Dichroism; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology; D019791:Guanidine / Q000494:pharmacology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011489:Protein Denaturation; D016914:Ribonuclease H / Q000737:chemistry*; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D013816:Thermodynamics; D016963:Thermus thermophilus / Q000201:enzymology
chemical_list: D016914:Ribonuclease H; D019791:Guanidine