Kinetic and thermodynamic allostery in the Ras protein family

Allostery, the transfer of information between distant parts of a macromolecule, is a fundamental feature of protein function and regulation. However, allosteric mechanisms are usually not explained by protein structure, requiring information on correlated fluctuations uniquely accessible to molecular simulation. Existing work to extract allosteric pathways from molecular dynamics simulations has focused on thermodynamic correlations. Here, we show how kinetic correlations encode complemen- tary information essential to explain observed variations in allosteric regulation. We applied kinetic and thermodynamic corre- lation analysis on atomistic simulations of H, K, and NRas isoforms in the apo, GTP, and GDP-bound states of Ras protein, with and without complexing to its downstream effector, Raf. We show that switch I and switch II are the primary components of thermodynamic and kinetic allosteric networks, consistent with the key roles of these two motifs. These networks connect the switches to an allosteric loop recently discovered from a crystal structure of HRas. This allosteric loop is inactive in KRas, but is coupled to the hydrolysis arm switch II in NRas and HRas. We find that the mechanism in the latter two isoforms are thermo- dynamic and kinetic, respectively. Binding of Raf-RBD further activates thermodynamic allostery in HRas and KRas but has limited effect on NRas. These results indicate that kinetic and thermodynamic correlations are both needed to explain protein function and allostery. These two distinct channels of allosteric regulation, and their combinatorial variability, may explain how subtle mutational differences can lead to diverse regulatory profiles among enzymatic proteins.SIGNIFICANCE Using the Ras family proteins as a model system, this work demonstrates that networks of kinetic correlations are needed, in addition to standard thermodynamic measures, to predict mutant and ligand-dependent allosteric regulation in enzymes.

Automated summary

Using the Ras family proteins as a model system, this study demonstrates that networks of kinetic correlations are needed, in addition to standard thermodynamic measures, to predict mutant and ligand-dependent allosteric regulation in enzymes.