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The molecular basis for protein kinase A anchoring revealed by solution NMR

Nature Structural Biology volume 6, pages 222–227 (1999)Cite this article

Abstract

Compartmentalization of signal transduction enzymes into signaling complexes is an important mechanism to ensure the specificity of intracellular events. Formation of these complexes is mediated by specialized protein motifs that participate in protein–protein interactions. The adenosine 3´,5´-cyclic monophosphate (cAMP)-dependent protein kinase (PKA) is localized through interaction of the regulatory (R) subunit dimer with A-kinase-anchoring proteins (AKAPs). We now report the solution structure of the type II PKA R-subunit fragment RIIα(1–44), which encompasses both the AKAP-binding and dimerization interfaces. This structure incorporates an X-type four-helix bundle dimerization motif with an extended hydrophobic face that is necessary for high-affinity AKAP binding. NMR data on the complex between RIIα(1–44) and an AKAP fragment reveals extensive contacts between the two proteins. Interestingly, this same dimerization motif is present in other signaling molecules, the S100 family. Therefore, the X-type four-helix bundle may represent a conserved fold for protein–protein interactions in signal transduction.

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Figure 1: Characterization of the RIIα(1–44) fragment.
Figure 2: Backbone fold and protomer orientation of RIIα(1–44).
Figure 3: Representations of the surface potential and hydrophobic core of RIIα(1–44).
Figure 4: Analysis of anchoring interactions.

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Acknowledgements

This work was supported in part by grants from the NIH to M.G.N., Z.E.H., V.C., J.D.S. and P.A.J., the American Heart Association to M.G.N., the American Cancer Society to P.A.J. and the Cancer Research Coordinating Center to P.A.J.. We gratefully acknowledge A. Deese, D. Eliezer, M. Foster, I. Radhakrishnan, R. Kriwacki, Nicholas Skelton, J. Chung and J. Love for helpful discussions; and J. Adams, A. Newton, G. Ghosh and S. Taylor for their critical reading of this manuscript.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, 92093-0359, California

    Marceen Glavic Newlon, Melinda Roy, Dimitrios Morikis & Patricia A. Jennings

  2. Howard Hughes Medical Institute, Vollum Institute, Portland, 97201-3098, Oregon

    Zachary E. Hausken & John D. Scott

  3. R.S. Dow Neurological Sciences Institute, Oregon Health Sciences University, Portland, 97209-1595, Oregon

    Vincent Coghlan

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  1. Marceen Glavic Newlon
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Correspondence to Patricia A. Jennings.

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Newlon, M., Roy, M., Morikis, D. et al. The molecular basis for protein kinase A anchoring revealed by solution NMR. Nat Struct Mol Biol 6, 222–227 (1999). https://doi.org/10.1038/6663

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