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Two PEST-like motifs regulate Ca2+/calpain-mediated cleavage of the CaVbeta3 subunit and provide important determinants for neuronal Ca2+ channel activity
- Source :
- European Journal of Neuroscience, European Journal of Neuroscience, Wiley, 2006, 23 (9), pp.2311-20. ⟨10.1111/j.1460-9568.2006.04749.x⟩, European Journal of Neuroscience, 2006, 23 (9), pp.2311-20. ⟨10.1111/j.1460-9568.2006.04749.x⟩
- Publication Year :
- 2006
- Publisher :
- HAL CCSD, 2006.
-
Abstract
- International audience; An increase in intracellular Ca2+ due to voltage-gated Ca2+ (CaV) channel opening represents an important trigger for a number of second-messenger-mediated effects ranging from neurotransmitter release to gene activation. Ca2+ entry occurs through the principal pore-forming protein but several ancillary subunits are known to more precisely tune ion influx. Among them, the CaVbeta subunits are perhaps the most important, given that they largely influence the biophysical and pharmacological properties of the channel. Notably, several functional features may be associated with specific structural regions of the CaVbeta subunits emphasizing the relevance of intramolecular domains in the physiology of these proteins. In the current report, we show that CaVbeta3 contains two PEST motifs and undergoes Ca2+ -dependent degradation which can be prevented by the specific calpain inhibitor calpeptin. Using mutant constructs lacking the PEST motifs, we present evidence that they are necessary for the cleavage of CaVbeta3 by calpain. Furthermore, the deletion of the PEST sequences did not affect the binding of CaVbeta3 to the ion-conducting CaV2.2 subunit and, when expressed in human embryonic kidney-293 cells, the PEST motif-deleted CaVbeta3 significantly increased whole-cell current density and retarded channel inactivation. Consistent with this observation, calpeptin treatment of human embryonic kidney-293 cells expressing wild-type CaVbeta3 resulted in an increase in current amplitude. Together, these findings suggest that calpain-mediated CaVbeta3 proteolysis may be an essential process for Ca2+ channel functional regulation.
- Subjects :
- Patch-Clamp Techniques
Time Factors
Protein Conformation
MAGUK proteins
HEK-293 cells
MESH: Calcium-Binding Proteins
Membrane Potentials
0302 clinical medicine
Protein structure
MESH: Protein Conformation
MESH: Dipeptides
Calcium-binding protein
Beta subunit
Calcium signaling
0303 health sciences
Voltage-dependent calcium channel
biology
General Neuroscience
Microfilament Proteins
MESH: Electric Stimulation
Calpain
Dipeptides
MESH: Protein Subunits
Cell biology
MESH: Calcium
MESH: Calcium Channels
Ca2+ channels
calpain
MESH: Mutation
Recombinant Fusion Proteins
Protein subunit
Blotting, Western
Transfection
MESH: Calcium Signaling
Article
Cell Line
03 medical and health sciences
MESH: Microfilament Proteins
MESH: Patch-Clamp Techniques
MESH: Dose-Response Relationship, Radiation
MESH: Recombinant Fusion Proteins
[SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology
Humans
Immunoprecipitation
MESH: Blotting, Western
MESH: Membrane Potentials
[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology
Calcium Signaling
Patch clamp
030304 developmental biology
MESH: Humans
MESH: Immunoprecipitation
MESH: Transfection
Calcium-Binding Proteins
HEK 293 cells
MESH: Time Factors
Dose-Response Relationship, Radiation
Electric Stimulation
MESH: Cell Line
PEST sequences
Protein Subunits
Mutation
biology.protein
Calcium
Calcium Channels
030217 neurology & neurosurgery
Subjects
Details
- Language :
- English
- ISSN :
- 0953816X and 14609568
- Database :
- OpenAIRE
- Journal :
- European Journal of Neuroscience, European Journal of Neuroscience, Wiley, 2006, 23 (9), pp.2311-20. ⟨10.1111/j.1460-9568.2006.04749.x⟩, European Journal of Neuroscience, 2006, 23 (9), pp.2311-20. ⟨10.1111/j.1460-9568.2006.04749.x⟩
- Accession number :
- edsair.doi.dedup.....ebc6d14a14edcd8d635f8c4a78a1879e