Multi-state recognition pathway of the intrinsically disordered protein kinase inhibitor by protein kinase A
Open Access
- 27 April 2020
- journal article
- research article
- Published by eLife Sciences Publications, Ltd in eLife
Abstract
In the nucleus, the spatiotemporal regulation of the catalytic subunit of cAMP-dependent protein kinase A (PKA-C) is orchestrated by an intrinsically disordered protein kinase inhibitor, PKI, which recruits the CRM1/RanGTP nuclear exporting complex. How the PKA-C/PKI complex assembles and recognizes CRM1/RanGTP is not well understood. Using NMR, SAXS, fluorescence, metadynamics, and Markov model analysis, we determined the multi-state recognition pathway for PKI. After a fast binding step in which PKA-C selects PKI's most competent conformations, PKI folds upon binding through a slow conformational rearrangement within the enzyme's binding pocket. The high-affinity and pseudo-substrate regions of PKI become more structured and the transient interactions with the kinase augment the helical content of the nuclear export sequence, which is then poised to recruit the CRM1/RanGTP complex for nuclear translocation. The multistate binding mechanism featured by PKA-C/PKI complex represents a paradigm on how disordered, ancillary proteins (or protein domains) are able to operate multiple functions such as inhibiting the kinase while recruiting other regulatory proteins for nuclear export.Funding Information
- National Institute of General Medical Sciences (GM 100310)
- National Institute of General Medical Sciences (GM 46736)
This publication has 120 references indexed in Scilit:
- Three-dimensional triple-resonance NMR Spectroscopy of isotopically enriched proteinsJournal of Magnetic Resonance, 2011
- Intrinsic disorder mediates the diverse regulatory functions of the Cdk inhibitor p21Nature Chemical Biology, 2011
- Dynamics connect substrate recognition to catalysis in protein kinase ANature Chemical Biology, 2010
- Theory, Practice, and Applications of Paramagnetic Relaxation Enhancement for the Characterization of Transient Low-Population States of Biological Macromolecules and Their ComplexesChemical Reviews, 2009
- Backbone NMR resonance assignment of the catalytic subunit of cAMP-dependent protein kinase A in complex with AMP-PNPBiomolecular NMR Assignments, 2009
- GROMACS 4: Algorithms for Highly Efficient, Load-Balanced, and Scalable Molecular SimulationJournal of Chemical Theory and Computation, 2008
- Allosteric cooperativity in protein kinase AProceedings of the National Academy of Sciences of the United States of America, 2008
- Practical aspects of 1H transverse paramagnetic relaxation enhancement measurements on macromoleculesJournal of Magnetic Resonance, 2007
- Measurement ofJand Dipolar Couplings from Simplified Two-Dimensional NMR SpectraJournal of Magnetic Resonance, 1998
- NMRPipe: A multidimensional spectral processing system based on UNIX pipesJournal of Biomolecular NMR, 1995