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. 1999 Oct 15;343(Pt 2):443–452.

Mechanism of A-kinase-anchoring protein 79 (AKAP79) and protein kinase C interaction.

M C Faux 1, E N Rollins 1, A S Edwards 1, L K Langeberg 1, A C Newton 1, J D Scott 1
PMCID: PMC1220573  PMID: 10510312

Abstract

The A-kinase-anchoring protein AKAP79 co-ordinates the location of cAMP-dependent protein kinase, phosphatase 2B (PP2B/calcineurin) and protein kinase C (PKC) at postsynaptic sites in neurons. In this report we focus on the mechanism of interaction between AKAP79 and PKC. We show that neither lipid activators nor kinase activation are required for association with AKAP79. The anchoring protein binds and inhibits the conserved catalytic core of PKCbetaII. AKAP79 also associates with conventional, novel and atypical isoforms of PKC in vitro and in vivo, and immunofluorescence staining of rat hippocampal neurons demonstrates that the murine anchoring-protein homologue AKAP150 is co-distributed with PKCalpha/beta, PKCepsilon or PKCiota. Binding of the AKAP79(31-52) peptide, which inhibits kinase activity, exposes the pseudosubstrate domain of PKCbetaII, allowing endoproteinase Arg-C proteolysis in the absence of kinase activators. Reciprocal experiments have identified two arginine residues at positions 39 and 40 that are essential for AKAP79(31-52) peptide inhibition of PKCbetaII. Likewise, the same mutations in the full-length anchoring protein reduced inhibition of PKCbetaII. Thus AKAP79 associates with multiple PKC isoforms through a mechanism involving protein-protein interactions at the catalytic core where binding of the anchoring protein inhibits kinase activity through displacement of the pseudosubstrate.

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Selected References

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  1. Behn-Krappa A., Newton A. C. The hydrophobic phosphorylation motif of conventional protein kinase C is regulated by autophosphorylation. Curr Biol. 1999 Jul 15;9(14):728–737. doi: 10.1016/s0960-9822(99)80332-7. [DOI] [PubMed] [Google Scholar]
  2. Carr D. W., Stofko-Hahn R. E., Fraser I. D., Cone R. D., Scott J. D. Localization of the cAMP-dependent protein kinase to the postsynaptic densities by A-kinase anchoring proteins. Characterization of AKAP 79. J Biol Chem. 1992 Aug 25;267(24):16816–16823. [PubMed] [Google Scholar]
  3. Chapline C., Mousseau B., Ramsay K., Duddy S., Li Y., Kiley S. C., Jaken S. Identification of a major protein kinase C-binding protein and substrate in rat embryo fibroblasts. Decreased expression in transformed cells. J Biol Chem. 1996 Mar 15;271(11):6417–6422. doi: 10.1074/jbc.271.11.6417. [DOI] [PubMed] [Google Scholar]
  4. Chapline C., Ramsay K., Klauck T., Jaken S. Interaction cloning of protein kinase C substrates. J Biol Chem. 1993 Apr 5;268(10):6858–6861. [PubMed] [Google Scholar]
  5. Chen L., Huang L. Y. Sustained potentiation of NMDA receptor-mediated glutamate responses through activation of protein kinase C by a mu opioid. Neuron. 1991 Aug;7(2):319–326. doi: 10.1016/0896-6273(91)90270-a. [DOI] [PubMed] [Google Scholar]
  6. Coghlan V. M., Perrino B. A., Howard M., Langeberg L. K., Hicks J. B., Gallatin W. M., Scott J. D. Association of protein kinase A and protein phosphatase 2B with a common anchoring protein. Science. 1995 Jan 6;267(5194):108–111. doi: 10.1126/science.7528941. [DOI] [PubMed] [Google Scholar]
  7. Dekker L. V., Parker P. J. Regulated binding of the protein kinase C substrate GAP-43 to the V0/C2 region of protein kinase C-delta. J Biol Chem. 1997 May 9;272(19):12747–12753. doi: 10.1074/jbc.272.19.12747. [DOI] [PubMed] [Google Scholar]
  8. Dell'Acqua M. L., Faux M. C., Thorburn J., Thorburn A., Scott J. D. Membrane-targeting sequences on AKAP79 bind phosphatidylinositol-4, 5-bisphosphate. EMBO J. 1998 Apr 15;17(8):2246–2260. doi: 10.1093/emboj/17.8.2246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dell'Acqua M. L., Scott J. D. Protein kinase A anchoring. J Biol Chem. 1997 May 16;272(20):12881–12884. doi: 10.1074/jbc.272.20.12881. [DOI] [PubMed] [Google Scholar]
  10. Edwards A. S., Newton A. C. Regulation of protein kinase C betaII by its C2 domain. Biochemistry. 1997 Dec 16;36(50):15615–15623. doi: 10.1021/bi9718752. [DOI] [PubMed] [Google Scholar]
  11. Faux M. C., Scott J. D. Molecular glue: kinase anchoring and scaffold proteins. Cell. 1996 Apr 5;85(1):9–12. doi: 10.1016/s0092-8674(00)81075-2. [DOI] [PubMed] [Google Scholar]
  12. Faux M. C., Scott J. D. More on target with protein phosphorylation: conferring specificity by location. Trends Biochem Sci. 1996 Aug;21(8):312–315. [PubMed] [Google Scholar]
  13. Faux M. C., Scott J. D. Regulation of the AKAP79-protein kinase C interaction by Ca2+/Calmodulin. J Biol Chem. 1997 Jul 4;272(27):17038–17044. doi: 10.1074/jbc.272.27.17038. [DOI] [PubMed] [Google Scholar]
  14. Fraser I. D., Tavalin S. J., Lester L. B., Langeberg L. K., Westphal A. M., Dean R. A., Marrion N. V., Scott J. D. A novel lipid-anchored A-kinase Anchoring Protein facilitates cAMP-responsive membrane events. EMBO J. 1998 Apr 15;17(8):2261–2272. doi: 10.1093/emboj/17.8.2261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gao T., Yatani A., Dell'Acqua M. L., Sako H., Green S. A., Dascal N., Scott J. D., Hosey M. M. cAMP-dependent regulation of cardiac L-type Ca2+ channels requires membrane targeting of PKA and phosphorylation of channel subunits. Neuron. 1997 Jul;19(1):185–196. doi: 10.1016/s0896-6273(00)80358-x. [DOI] [PubMed] [Google Scholar]
  16. Glantz S. B., Amat J. A., Rubin C. S. cAMP signaling in neurons: patterns of neuronal expression and intracellular localization for a novel protein, AKAP 150, that anchors the regulatory subunit of cAMP-dependent protein kinase II beta. Mol Biol Cell. 1992 Nov;3(11):1215–1228. doi: 10.1091/mbc.3.11.1215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Glantz S. B., Li Y., Rubin C. S. Characterization of distinct tethering and intracellular targeting domains in AKAP75, a protein that links cAMP-dependent protein kinase II beta to the cytoskeleton. J Biol Chem. 1993 Jun 15;268(17):12796–12804. [PubMed] [Google Scholar]
  18. House C., Kemp B. E. Protein kinase C contains a pseudosubstrate prototope in its regulatory domain. Science. 1987 Dec 18;238(4834):1726–1728. doi: 10.1126/science.3686012. [DOI] [PubMed] [Google Scholar]
  19. Hubbard M. J., Cohen P. On target with a new mechanism for the regulation of protein phosphorylation. Trends Biochem Sci. 1993 May;18(5):172–177. doi: 10.1016/0968-0004(93)90109-z. [DOI] [PubMed] [Google Scholar]
  20. Hunter T. Protein kinases and phosphatases: the yin and yang of protein phosphorylation and signaling. Cell. 1995 Jan 27;80(2):225–236. doi: 10.1016/0092-8674(95)90405-0. [DOI] [PubMed] [Google Scholar]
  21. Hyatt S. L., Klauck T., Jaken S. Protein kinase C is localized in focal contacts of normal but not transformed fibroblasts. Mol Carcinog. 1990;3(2):45–53. doi: 10.1002/mc.2940030202. [DOI] [PubMed] [Google Scholar]
  22. Kemp B. E., Pearson R. B. Protein kinase recognition sequence motifs. Trends Biochem Sci. 1990 Sep;15(9):342–346. doi: 10.1016/0968-0004(90)90073-k. [DOI] [PubMed] [Google Scholar]
  23. Keranen L. M., Dutil E. M., Newton A. C. Protein kinase C is regulated in vivo by three functionally distinct phosphorylations. Curr Biol. 1995 Dec 1;5(12):1394–1403. doi: 10.1016/s0960-9822(95)00277-6. [DOI] [PubMed] [Google Scholar]
  24. Klauck T. M., Faux M. C., Labudda K., Langeberg L. K., Jaken S., Scott J. D. Coordination of three signaling enzymes by AKAP79, a mammalian scaffold protein. Science. 1996 Mar 15;271(5255):1589–1592. doi: 10.1126/science.271.5255.1589. [DOI] [PubMed] [Google Scholar]
  25. Klauck T. M., Scott J. D. The postsynaptic density: a subcellular anchor for signal transduction enzymes. Cell Signal. 1995 Nov;7(8):747–757. doi: 10.1016/0898-6568(95)02003-9. [DOI] [PubMed] [Google Scholar]
  26. Mochly-Rosen D. Localization of protein kinases by anchoring proteins: a theme in signal transduction. Science. 1995 Apr 14;268(5208):247–251. doi: 10.1126/science.7716516. [DOI] [PubMed] [Google Scholar]
  27. Newton A. C. Protein kinase C: structure, function, and regulation. J Biol Chem. 1995 Dec 1;270(48):28495–28498. doi: 10.1074/jbc.270.48.28495. [DOI] [PubMed] [Google Scholar]
  28. Orr J. W., Keranen L. M., Newton A. C. Reversible exposure of the pseudosubstrate domain of protein kinase C by phosphatidylserine and diacylglycerol. J Biol Chem. 1992 Aug 5;267(22):15263–15266. [PubMed] [Google Scholar]
  29. Orr J. W., Newton A. C. Interaction of protein kinase C with phosphatidylserine. 2. Specificity and regulation. Biochemistry. 1992 May 19;31(19):4667–4673. doi: 10.1021/bi00134a019. [DOI] [PubMed] [Google Scholar]
  30. Orr J. W., Newton A. C. Intrapeptide regulation of protein kinase C. J Biol Chem. 1994 Mar 18;269(11):8383–8387. [PubMed] [Google Scholar]
  31. Orr J. W., Newton A. C. Requirement for negative charge on "activation loop" of protein kinase C. J Biol Chem. 1994 Nov 4;269(44):27715–27718. [PubMed] [Google Scholar]
  32. Pawson T., Scott J. D. Signaling through scaffold, anchoring, and adaptor proteins. Science. 1997 Dec 19;278(5346):2075–2080. doi: 10.1126/science.278.5346.2075. [DOI] [PubMed] [Google Scholar]
  33. Ron D., Chen C. H., Caldwell J., Jamieson L., Orr E., Mochly-Rosen D. Cloning of an intracellular receptor for protein kinase C: a homolog of the beta subunit of G proteins. Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):839–843. doi: 10.1073/pnas.91.3.839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ron D., Mochly-Rosen D. An autoregulatory region in protein kinase C: the pseudoanchoring site. Proc Natl Acad Sci U S A. 1995 Jan 17;92(2):492–496. doi: 10.1073/pnas.92.2.492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rosenmund C., Carr D. W., Bergeson S. E., Nilaver G., Scott J. D., Westbrook G. L. Anchoring of protein kinase A is required for modulation of AMPA/kainate receptors on hippocampal neurons. Nature. 1994 Apr 28;368(6474):853–856. doi: 10.1038/368853a0. [DOI] [PubMed] [Google Scholar]
  36. Rubin C. S. A kinase anchor proteins and the intracellular targeting of signals carried by cyclic AMP. Biochim Biophys Acta. 1994 Dec 30;1224(3):467–479. [PubMed] [Google Scholar]
  37. Staudinger J., Lu J., Olson E. N. Specific interaction of the PDZ domain protein PICK1 with the COOH terminus of protein kinase C-alpha. J Biol Chem. 1997 Dec 19;272(51):32019–32024. doi: 10.1074/jbc.272.51.32019. [DOI] [PubMed] [Google Scholar]
  38. Staudinger J., Zhou J., Burgess R., Elledge S. J., Olson E. N. PICK1: a perinuclear binding protein and substrate for protein kinase C isolated by the yeast two-hybrid system. J Cell Biol. 1995 Feb;128(3):263–271. doi: 10.1083/jcb.128.3.263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tong G., Shepherd D., Jahr C. E. Synaptic desensitization of NMDA receptors by calcineurin. Science. 1995 Mar 10;267(5203):1510–1512. doi: 10.1126/science.7878472. [DOI] [PubMed] [Google Scholar]
  40. Wang L. Y., Dudek E. M., Browning M. D., MacDonald J. F. Modulation of AMPA/kainate receptors in cultured murine hippocampal neurones by protein kinase C. J Physiol. 1994 Mar 15;475(3):431–437. doi: 10.1113/jphysiol.1994.sp020083. [DOI] [PMC free article] [PubMed] [Google Scholar]

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