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Structural determinants of calmodulin binding to the intracellular C-terminal domain of the metabotropic glutamate receptor 7A

Structural determinants of calmodulin binding to the intracellular C-terminal domain of the metabotropic glutamate receptor 7A
Structural determinants of calmodulin binding to the intracellular C-terminal domain of the metabotropic glutamate receptor 7A
Calmodulin (CaM) binds in a Ca2+-dependent manner to the intracellular C-terminal domains of most group III metabotropic glutamate receptors (mGluRs). Here we combined mutational and biophysical approaches to define the structural basis of CaM binding to mGluR 7A. Ca2+/CaM was found to interact with mGluR 7A primarily via its C-lobe at a 1:1 CaM:C-tail stoichiometry. Pulldown experiments with mutant CaM and mGluR 7A C-tail constructs and high resolution NMR with peptides corresponding to the CaM binding region of mGluR 7A allowed us to define hydrophobic and ionic interactions required for Ca2+/CaM binding and identified a 1-8-14 CaM-binding motif. The Ca2+/CaM·mGluR 7A peptide complex displays a classical wraparound structure that closely resembles that formed by Ca2+/CaM upon binding to smooth muscle myosin light chain kinase. Our data provide insight into how Ca2+/CaM regulates group III mGluR signaling via competition with intracellular proteins for receptor-binding sites.
0021-9258
5577-5588
Scheschonka, Astrid
a9508213-1021-42fc-903f-ac555b5a9016
Findlow, Stuart C.
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Schemm, Rudolf
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El Far, Oussama
d8170148-c0d9-447e-8366-ccdca6f6129a
Caldwell, John H.
eb81be92-ecaa-4f40-b19e-f5232c79f997
Crump, Matthew P.
ed31b5fd-23f6-434c-a38c-da3cb7b27402
Holden-Dye, Kate
23de5b43-7e0e-4704-a46c-b392781822c7
O'Connor, Vincent
8021b06c-01a0-4925-9dde-a61c8fe278ca
Betz, Heinrich
1833229b-589c-4f4b-b10c-cd40f68977a5
Werner, Jorn M.
1b02513a-8310-4f4f-adac-dc2a466bd115
Scheschonka, Astrid
a9508213-1021-42fc-903f-ac555b5a9016
Findlow, Stuart C.
0a7193ad-45cf-4613-a35e-3fd019a92ebe
Schemm, Rudolf
d94f70e8-1582-417b-a136-8ddc2e743070
El Far, Oussama
d8170148-c0d9-447e-8366-ccdca6f6129a
Caldwell, John H.
eb81be92-ecaa-4f40-b19e-f5232c79f997
Crump, Matthew P.
ed31b5fd-23f6-434c-a38c-da3cb7b27402
Holden-Dye, Kate
23de5b43-7e0e-4704-a46c-b392781822c7
O'Connor, Vincent
8021b06c-01a0-4925-9dde-a61c8fe278ca
Betz, Heinrich
1833229b-589c-4f4b-b10c-cd40f68977a5
Werner, Jorn M.
1b02513a-8310-4f4f-adac-dc2a466bd115

Scheschonka, Astrid, Findlow, Stuart C., Schemm, Rudolf, El Far, Oussama, Caldwell, John H., Crump, Matthew P., Holden-Dye, Kate, O'Connor, Vincent, Betz, Heinrich and Werner, Jorn M. (2008) Structural determinants of calmodulin binding to the intracellular C-terminal domain of the metabotropic glutamate receptor 7A. The Journal of Biological Chemistry, 283 (9), 5577-5588. (doi:10.1074/jbc.M709505200). (PMID:18089570)

Record type: Article

Abstract

Calmodulin (CaM) binds in a Ca2+-dependent manner to the intracellular C-terminal domains of most group III metabotropic glutamate receptors (mGluRs). Here we combined mutational and biophysical approaches to define the structural basis of CaM binding to mGluR 7A. Ca2+/CaM was found to interact with mGluR 7A primarily via its C-lobe at a 1:1 CaM:C-tail stoichiometry. Pulldown experiments with mutant CaM and mGluR 7A C-tail constructs and high resolution NMR with peptides corresponding to the CaM binding region of mGluR 7A allowed us to define hydrophobic and ionic interactions required for Ca2+/CaM binding and identified a 1-8-14 CaM-binding motif. The Ca2+/CaM·mGluR 7A peptide complex displays a classical wraparound structure that closely resembles that formed by Ca2+/CaM upon binding to smooth muscle myosin light chain kinase. Our data provide insight into how Ca2+/CaM regulates group III mGluR signaling via competition with intracellular proteins for receptor-binding sites.

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e-pub ahead of print date: 18 December 2007
Published date: February 2008
Organisations: Centre for Biological Sciences

Identifiers

Local EPrints ID: 143015
URI: http://eprints.soton.ac.uk/id/eprint/143015
ISSN: 0021-9258
PURE UUID: 3e786fd2-fb8a-47a3-87d2-c734c24bd4ce
ORCID for Vincent O'Connor: ORCID iD orcid.org/0000-0003-3185-5709
ORCID for Jorn M. Werner: ORCID iD orcid.org/0000-0002-4712-1833

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Date deposited: 08 Apr 2010 09:04
Last modified: 14 Mar 2024 02:48

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Contributors

Author: Astrid Scheschonka
Author: Stuart C. Findlow
Author: Rudolf Schemm
Author: Oussama El Far
Author: John H. Caldwell
Author: Matthew P. Crump
Author: Kate Holden-Dye
Author: Heinrich Betz
Author: Jorn M. Werner ORCID iD

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