Thermostability of Enzymes from Molecular Dynamics Simulations.
Thermodynamic stability is a central requirement for protein function, and one goal of protein engineering is improvement of stability, particularly for applications in biotechnology. Herein, molecular dynamics simulations are used to predict in vitro thermostability of members of the bacterial ribonuclease HI (RNase H) family of endonucleases. The temperature dependence of the generalized order parameter, S, for four RNase H homologues, from psychrotrophic, mesophilic, and thermophilic organisms, is highly correlated with experimentally determined melting temperatures and with calculated free energies of folding at the midpoint temperature of the simulations. This study provides an approach for in silico mutational screens to improve thermostability of biologically and industrially relevant enzymes.
Department of Biochemistry and Molecular Biophysics, Columbia University , New York, New York 10032, United States.
Tim
T
Zeiske
Department of Biochemistry and Molecular Biophysics, Columbia University , New York, New York 10032, United States.
Kate A
KA
Stafford
Department of Biochemistry and Molecular Biophysics, Columbia University , New York, New York 10032, United States.
Arthur G
AG
Palmer
D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D056004:Molecular Dynamics Simulation; D017510:Protein Folding; D055550:Protein Stability; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D016914:Ribonuclease H / Q000737:chemistry* / Q000378:metabolism; D020592:Shewanella / Q000201:enzymology; D016963:Thermus thermophilus / Q000201:enzymology; D044366:Transition Temperature
D001426:Bacterial Proteins; D016914:Ribonuclease H; C078856:ribonuclease HI