TY - JOUR TI - A dynamic mechanism for allosteric activation of Aurora kinase A by activation loop phosphorylation AU - Ruff, Emily F AU - Muretta, Joseph M AU - Thompson, Andrew R AU - Lake, Eric W AU - Cyphers, Soreen AU - Albanese, Steven K AU - Hanson, Sonya M AU - Behr, Julie M AU - Thomas, David D AU - Chodera, John D AU - Levinson, Nicholas M A2 - Weis, William I VL - 7 PY - 2018 DA - 2018/02/21 SP - e32766 C1 - eLife 2018;7:e32766 DO - 10.7554/eLife.32766 UR - https://doi.org/10.7554/eLife.32766 AB - Many eukaryotic protein kinases are activated by phosphorylation on a specific conserved residue in the regulatory activation loop, a post-translational modification thought to stabilize the active DFG-In state of the catalytic domain. Here we use a battery of spectroscopic methods that track different catalytic elements of the kinase domain to show that the ~100 fold activation of the mitotic kinase Aurora A (AurA) by phosphorylation occurs without a population shift from the DFG-Out to the DFG-In state, and that the activation loop of the activated kinase remains highly dynamic. Instead, molecular dynamics simulations and electron paramagnetic resonance experiments show that phosphorylation triggers a switch within the DFG-In subpopulation from an autoinhibited DFG-In substate to an active DFG-In substate, leading to catalytic activation. This mechanism raises new questions about the functional role of the DFG-Out state in protein kinases. KW - protein kinase KW - phosphorylation KW - protein dynamics JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -