The A-kinase Anchoring Domain of Type IIα cAMP-dependent Protein Kinase Is Highly Helical*
- Marceen Glavic Newlon‡,
- Melinda Roy‡,
- Zachary E. Hausken§,
- John D. Scott§ and
- Patricia A. Jennings‡¶
- From the ‡Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, California 92093-0359 and the§Howard Hughes Medical Institute, Vollum Institute, Portland, Oregon 97201-3098
Abstract
Subcellular localization of the type II cAMP-dependent protein kinase is controlled by interaction of the regulatory subunit with A-KinaseAnchoring Proteins (AKAPs). This contribution examines the solution structure of a 44-residue region that is sufficient for high affinity binding to AKAPs. The N-terminal dimerization domain of the type IIα regulatory subunit of cAMP-dependent protein kinase was expressed to high levels on minimal media and uniformly isotopically enriched with15N and 13C nuclei. Sequence-specific backbone and side chain resonance assignments have been made for greater than 95% of the amino acids in the free dimerization domain using high resolution multidimensional heteronuclear NMR techniques. Contrary to the results from secondary structure prediction algorithms, our analysis indicates that the domain is highly helical with a single 3–5-residue sequence involved in a β-strand. The assignments and secondary structure analysis provide the basis for analyzing the structure and dynamics of the dimerization domain both free and complexed with specific anchoring proteins.
Footnotes
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↵* This work was supported by National Institutes of Health Grants GM07313 (to M. G. N.), CA09523 (to M. R.), and DK44239 (to J. D. S. and Z. E. H.) and the Cancer Research Coordinating Committee (to P. A. J.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵¶ To whom correspondence should be addressed.
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↵1 The abbreviations used are: AKAP, protein kinase A anchoring protein; PKA, cAMP-dependent protein kinase; R2 subunit, cAMP-dependent protein kinase regulatory subunit dimer; C subunit, cAMP-dependent protein kinase catalytic subunit; TOCSY, total correlation spectroscopy; NOE nuclear Overhauser effect; NOESY, nuclear Overhauser enhancement spectroscopy; CSI, chemical shift index; HSQC, heteronuclear single quantum coherence; HMQC, heteronuclear multiple quantum coherence.
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↵2 M. Newlon and P. A. Jennings, manuscript in preparation.
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- Received June 4, 1997.
- Revision received July 3, 1997.
