| RRC ID |
62064
|
| 著者 |
Alli AA, Gower WR Jr.
|
| タイトル |
The C type natriuretic peptide receptor tethers AHNAK1 at the plasma membrane to potentiate arachidonic acid-induced calcium mobilization.
|
| ジャーナル |
Am J Physiol Cell Physiol
|
| Abstract |
Arachidonic acid (AA) liberated from membrane phospholipids is known to activate phospholipase C gamma1 (PLCgamma1) concurrently with AHNAK in nonneuronal cells. The recruitment of AHNAK from the nucleus is required for it to activate PLCgamma1 at the plasma membrane. Here, we identify the C-type natriuretic peptide receptor (NPR-C), an atypical G protein-coupled receptor, as a protein binding partner for AHNAK1 in various cell types. Mass spectrometry and MASCOT analysis of excised bands from NPR-C immunoprecipitation studies revealed multiple signature peptides corresponding to AHNAK1. Glutathione S-transferase (GST) pulldown assays using GST- AHNAK1 fusion proteins corresponding to each of the distinct domains of AHNAK1 showed the C1 domain of AHNAK1 associates with NPR-C. The role of NPR-C in mediating AA-dependent AHNAK1 calcium signaling was explored in various cell types, including 3T3-L1 preadipocytes during the early stages of differentiation. Sucrose density gradient centrifugation studies showed AHNAK1 resides in the nucleus, cytoplasm, and at the plasma membrane, but small interfering RNA (siRNA)-mediated knockdown of NPR-C resulted in AHNAK1 accumulation in the nucleus. Overexpression of a portion of AHNAK1 resulted in augmentation of intracellular calcium mobilization, whereas siRNA-mediated knockdown of NPR-C or AHNAK1 protein resulted in attenuation of intracellular calcium mobilization in response to phorbol 12-myristate 13-acetate. We characterize the novel association between AHNAK1 and NPR-C and provide evidence that this association potentiates the AA-induced mobilization of intracellular calcium. We address the role of intracellular calcium in the various cell types that AHNAK1 and NPR-C were found to associate.
|
| 巻・号 |
297(5)
|
| ページ |
C1157-67
|
| 公開日 |
2009-11-1
|
| DOI |
10.1152/ajpcell.00219.2009
|
| PII |
00219.2009
|
| PMID |
19710363
|
| MeSH |
Animals
Arachidonic Acid / metabolism*
Blotting, Western
Calcium Signaling / physiology*
Cell Line
Cell Membrane / metabolism*
Electrophoresis, Polyacrylamide Gel
Gene Expression
HeLa Cells
Humans
Immunoprecipitation
Membrane Proteins / metabolism*
Mice
Microscopy, Confocal
Muscle, Smooth, Vascular / metabolism
Neoplasm Proteins / metabolism*
Rats
Receptors, Atrial Natriuretic Factor / metabolism*
|
| IF |
3.485
|
| リソース情報 |
| ヒト・動物細胞 |
RGM1(RCB0876) |
