Journal of Biological Chemistry
Volume 276, Issue 43, 26 October 2001, Pages 40104-40112
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MECHANISMS OF SIGNAL TRANSDUCTION
Synaptic Scaffolding Proteins in Rat Brain: ANKYRIN REPEATS OF THE MULTIDOMAIN Shank PROTEIN FAMILY INTERACT WITH THE CYTOSKELETAL PROTEIN α-FODRIN*

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The postsynaptic density is the ultrastructural entity containing the neurotransmitter reception apparatus of excitatory synapses in the brain. A recently identified family of multidomain proteins termed Src homology 3 domain and ankyrin repeat-containing (Shank), also known as proline-rich synapse-associated protein/somatostatin receptor-interacting protein, plays a central role in organizing the subsynaptic scaffold by interacting with several synaptic proteins including the glutamate receptors. We used the N-terminal ankyrin repeats of Shank1 and -3 to search for interacting proteins by yeast two-hybrid screening and by affinity chromatography. By cDNA sequencing and mass spectrometry the cytoskeletal protein α-fodrin was identified as an interacting molecule. The interaction was verified by pull-down assays and by coimmunoprecipitation experiments from transfected cells and brain extracts. Mapping of the interacting domains of α-fodrin revealed that the highly conserved spectrin repeat 21 is sufficient to bind to the ankyrin repeats. Both interacting partners are coexpressed widely in the rat brain and are colocalized in synapses of hippocampal cultures. Our data indicate that the Shank1 and -3 family members provide multiple independent connections between synaptic glutamate receptor complexes and the cytoskeleton.

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Published, JBC Papers in Press, August 16, 2001, DOI 10.1074/jbc.M102454200

*

This work was supported in part by Deutsche Forschungsgemeinschaft Grants SFB545/B7 (to H.-J. K. and D. R.), SFB426/A1 (to E. D. G.), and KR1879/2–1 (to M. R. K.), European Commission Grant QLG3-CT-1999-00908 (to D. R.), Fonds der Chemischen Industrie (to E. D. G and C. W.), and Universität Münster IMF and IZKF/F1 (to T. M. B.).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.

Work presented as part of a thesis.

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These authors contributed equally to this work.