Apo-calmodulin binds with its C-terminal domain to the N-methyl-D-aspartate receptor NR1 C0 region

Zeynep Akyol; Jason A Bartos; Michelle A Merrill; Laurel A Faga; Olav R Jaren; Madeline A Shea; Johannes W Hell

doi:10.1074/jbc.M302542200

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Apo-calmodulin binds with its C-terminal domain to the N-methyl-D-aspartate receptor NR1 C0 region

Journal article

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Peer reviewed

Apo-calmodulin binds with its C-terminal domain to the N-methyl-D-aspartate receptor NR1 C0 region

Zeynep Akyol, Jason A Bartos, Michelle A Merrill, Laurel A Faga, Olav R Jaren, Madeline A Shea and Johannes W Hell

The Journal of biological chemistry, Vol.279(3), pp.2166-2175

01/16/2004

DOI: 10.1074/jbc.M302542200

PMID: 14530275

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Abstract

Calmodulin (CaM) is the major Ca2+ sensor in eukaryotic cells. It consists of four EF-hand Ca2+ binding motifs, two in its N-terminal domain and two in its C-terminal domain. Through a negative feedback loop, CaM inhibits Ca2+ influx through N-methyl-D-aspartate-type glutamate receptors in neurons by binding to the C0 region in the cytosolic tail of the NR1 subunit. Ca2+ -depleted (apo)CaM is pre-associated with a variety of ion channels for fast and effective regulation of channel activities upon Ca2+ influx. Using the NR1 C0 region for fluorescence and circular dichroism spectroscopy studies we found that not only Ca2+ -saturated CaM but also apoCaM bound to NR1 C0. In vitro interaction assays showed that apoCaM also binds specifically to full-length NR1 solubilized from rat brain and to the complete C terminus of the NR1 splice form that contains the C0 plus C2' domain. The Ca2+ -independent interaction of CaM was also observed with the isolated C-but not N-terminal fragment of calmodulin in the independent spectroscopic assays. Fluorescence polarization studies indicated that apoCaM associated via its C-terminal domain with NR1 C0 in an extended conformation and collapsed to adopt a more compact conformation of faster rotational mobility in its complex with NR1 C0 upon addition of Ca2+. Our results indicate that apoCaM is associated with NR1 and that the complex of CaM bound to NR1 C0 undergoes a dramatic conformational change when Ca2+ binds to CaM.

Calmodulin - metabolism

Calcium - metabolism

Receptors, N-Methyl-D-Aspartate - chemistry

Receptors, N-Methyl-D-Aspartate - metabolism

Spectrometry, Fluorescence

Calmodulin - chemistry

Fluorescence Polarization

Circular Dichroism

Details

Title: Subtitle: Apo-calmodulin binds with its C-terminal domain to the N-methyl-D-aspartate receptor NR1 C0 region
Creators: Zeynep Akyol - Department of Biochemistry, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA 52242-1109, USA
Jason A Bartos
Michelle A Merrill
Laurel A Faga
Olav R Jaren
Madeline A Shea
Johannes W Hell
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.279(3), pp.2166-2175
DOI: 10.1074/jbc.M302542200
PMID: 14530275
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: R01 NS 35563 / NINDS NIH HHS R01 GM 57001 / NIGMS NIH HHS T32 GM007337 / NIGMS NIH HHS
Language: English
Date published: 01/16/2004
Academic Unit: Molecular Physiology and Biophysics; Iowa Neuroscience Institute; Biochemistry and Molecular Biology
Record Identifier: 9984024404502771

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