title: Regulation of Ras Paralog Thermostability by Networks of Buried Ionizable Groups.
abstract: Protein folding is governed by a variety of molecular forces including hydrophobic and ionic interactions. Less is known about the molecular determinants of protein stability. Here we used a recently developed computer algorithm (pHinder) to investigate the relationship between buried charge and thermostability. Our analysis revealed that charge networks in the protein core are generally smaller in thermophilic organisms as compared to mesophilic organisms. To experimentally test whether core network size influences protein thermostability, we purified 18 paralogous Ras superfamily GTPases from yeast and determined their melting temperatures (Tm, or temperature at which 50% of the protein is unfolded). This analysis revealed a wide range of Tm values (35-63 °C) that correlated significantly (R = 0.87) with core network size. These results suggest that thermostability depends in part on the arrangement of ionizable side chains within a protein core. An improved capacity to predict protein thermostability may be useful for selecting the best candidates for protein crystallography, the development of protein-based therapeutics, as well as for industrial enzyme applications.
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina 27599, United States.
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Isom
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina 27599, United States.
authors.forename: Vishwajith
authors.initials: V
authors.lastname: Sridharan
authors.affiliation: Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill , Chapel Hill, North Carolina 27599, United States.
authors.forename: Henrik G
authors.initials: HG
authors.lastname: Dohlman
mesh_terms: D003545:Cysteine / Q000737:chemistry; D004795:Enzyme Stability; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D011489:Protein Denaturation; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*; D018631:ras Proteins / Q000737:chemistry*
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D018631:ras Proteins; D003545:Cysteine