Abstract
To understand the molecular mechanism underlying phosphoryl transfer of cAMP-dependent protein kinase, the structure of the catalytic subunit in complex with ADP, aluminum fluoride, Mg2+ ions and a substrate peptide was determined at 2.0 Å resolution. Aluminum fluoride was modeled as AlF3 in a planar geometry; it is positioned 2.3 Å from both the donor oxygen of ADP and the hydroxyl group of the recipient Ser residue. In this configuration, the aluminum atom forms a trigonal bipyramidal coordination with the oxygen atoms of the donor and recipient groups at the apical positions. This arrangement suggests that aluminum fluoride mimics the transition state and provides the first direct structural evidence for the in-line mechanism of phosphoryl transfer in a protein kinase.
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Acknowledgements
We thank D. Johnson for his valuable comments, E. Radzio-Andzelm for generating Fig. 2, C. Juliano for the protein preparation and N. Nguyen for X-ray data collection support. Portions of this research were carried out at the Stanford Synchrotron Radiation Laboratory, a national user facility operated by Stanford University on behalf of the U.S. Department of Energy, Office of Basic Energy Sciences. The SSRL Structural Molecular Biology Program is supported by the Department of Energy, Office of Biological and Environmental Research; the National Institutes of Health, National Center for Research Resources, Biomedical Technology Program; and the National Institute of General Medical Sciences. This work was supported by a grant from the National Institutes of Health (to S.S.T.). P.A. was supported by an NIH training grant.
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Madhusudan, Akamine, P., Xuong, NH. et al. Crystal structure of a transition state mimic of the catalytic subunit of cAMP-dependent protein kinase. Nat Struct Mol Biol 9, 273–277 (2002). https://doi.org/10.1038/nsb780
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DOI: https://doi.org/10.1038/nsb780
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