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Distinct enzyme combinations in AKAP signalling complexes permit functional diversity

Nature Cell Biology volume 7, pages 1066–1073 (2005)Cite this article

Abstract

Specificity in cell signalling can be influenced by the targeting of different enzyme combinations to substrates. The A-kinase anchoring protein AKAP79/150 is a multivalent scaffolding protein that coordinates the subcellular localization of second-messenger-regulated enzymes, such as protein kinase A, protein kinase C and protein phosphatase 2B. We developed a new strategy that combines RNA interference of the endogenous protein with a protocol that selects cells that have been rescued with AKAP79/150 forms that are unable to anchor selected enzymes. Using this approach, we show that AKAP79/150 coordinates different enzyme combinations to modulate the activity of two distinct neuronal ion channels: AMPA-type glutamate receptors and M-type potassium channels. Utilization of distinct enzyme combinations in this manner provides a means to expand the repertoire of cellular events that the same AKAP modulates.

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Figure 1: RNA interference of AKAP79 in HEK293 cells.
Figure 2: PP2B is a fundamental component of the GluR1–AKAP79/150 signalling network in HEK293 cells.
Figure 3: RNA interference of SAP97 in HEK293 cells.
Figure 4: Modulation of AMPA channels with modified AKAP complexes.
Figure 5: PKA activation is required for maintaining AMPA currents.
Figure 6: AKAP150 facilitates selective M-current suppression.
Figure 7: Functional dissection of an M-channel–AKAP complex.

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Acknowledgements

We thank J. J. Carlisle Michel and R. Goodman for insightful comments on the manuscript and R. Mouton for technical assistance. This work was supported by grant no. GM48231 from the National Institutes of Health to J.D.S.

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Authors and Affiliations

  1. Howard Hughes Medical Institute, Vollum Institute, Oregon Health and Science University, 3181 S.W. Sam Jackson Park Road, Portland, OR 97239, USA

    Naoto Hoshi, Lorene K. Langeberg & John D. Scott

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  1. Naoto Hoshi
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Correspondence to John D. Scott.

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Supplementary figures S1, S2, S3 and S4 (PDF 419 kb)

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Hoshi, N., Langeberg, L. & Scott, J. Distinct enzyme combinations in AKAP signalling complexes permit functional diversity. Nat Cell Biol 7, 1066–1073 (2005). https://doi.org/10.1038/ncb1315

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