A dynamic mechanism for allosteric activation of Aurora kinase A by activation loop phosphorylation
Abstract
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.
Article and author information
Author details
Funding
National Institutes of Health (R00 Award GM102288)
- Nicholas M Levinson
National Institutes of Health (R21 Award CA217695)
- Nicholas M Levinson
National Institutes of Health (NRSA Award F32GM120817)
- Emily F Ruff
National Institutes of Health (P30-CA008748)
- John D Chodera
National Institutes of Health (GM121505)
- John D Chodera
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2018, Ruff et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
Metrics
-
- 5,113
- views
-
- 729
- downloads
-
- 62
- citations
Views, downloads and citations are aggregated across all versions of this paper published by eLife.