TY - JOUR
T1 - Critical role of substrate conformational change in the proton transfer process catalyzed by 4-oxalocrotonate tautomerase
AU - Ruiz-Pernía, J. Javier
AU - Garcia-Viloca, Mireia
AU - Bhattacharyya, Sudeep
AU - Gao, Jiali
AU - Truhlar, Donald G.
AU - Tuñón, Iñaki
PY - 2009/2/25
Y1 - 2009/2/25
N2 - 4-Oxalocrotonate tautomerase enzyme (4-OT) catalyzes the isomerization of 2-oxo-4-hexenedioate to 2-oxo-3-hexenedioate. The chemical process involves two proton transfers, one from a carbon of the substrate to the nitrogen of Pro1 and another from this nitrogen atom to a different carbon of the substrate. In this paper the isomerization has been studied using the combined quantum mechanical and molecular mechanical method with a dual-level treatment of the quantum subsystem employing the MPW1BK density functional as the higher level. Exploration of the potential energy surface shows that the process is stepwise, with a stable intermediate state corresponding to the deprotonated substrate and a protonated proline. The rate constant of the overall process has been evaluated using ensemble-averaged variational transition state theory, including the quantized vibrational motion of a primary zone of active-site atoms and a transmission coefficient based on an ensemble of optimized reaction coordinates to account for recrossing trajectories and optimized multidimensional tunneling. The two proton-transfer steps have similar free energy barriers, but the transition state associated with the first proton transfer is found to be higher in energy. The calculations show that reaction progress is coupled to a conformational change of the substrate, so it is important that the simulation allows this flexibility. The coupled conformational change is promoted by changes in the electron distribution of the substrate that take place as the proton transfers occur. © 2009 American Chemical Society.
AB - 4-Oxalocrotonate tautomerase enzyme (4-OT) catalyzes the isomerization of 2-oxo-4-hexenedioate to 2-oxo-3-hexenedioate. The chemical process involves two proton transfers, one from a carbon of the substrate to the nitrogen of Pro1 and another from this nitrogen atom to a different carbon of the substrate. In this paper the isomerization has been studied using the combined quantum mechanical and molecular mechanical method with a dual-level treatment of the quantum subsystem employing the MPW1BK density functional as the higher level. Exploration of the potential energy surface shows that the process is stepwise, with a stable intermediate state corresponding to the deprotonated substrate and a protonated proline. The rate constant of the overall process has been evaluated using ensemble-averaged variational transition state theory, including the quantized vibrational motion of a primary zone of active-site atoms and a transmission coefficient based on an ensemble of optimized reaction coordinates to account for recrossing trajectories and optimized multidimensional tunneling. The two proton-transfer steps have similar free energy barriers, but the transition state associated with the first proton transfer is found to be higher in energy. The calculations show that reaction progress is coupled to a conformational change of the substrate, so it is important that the simulation allows this flexibility. The coupled conformational change is promoted by changes in the electron distribution of the substrate that take place as the proton transfers occur. © 2009 American Chemical Society.
U2 - 10.1021/ja8087423
DO - 10.1021/ja8087423
M3 - Article
SN - 0002-7863
VL - 131
SP - 2687
EP - 2698
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 7
ER -