| RRC ID |
18561
|
| 著者 |
Okano I, Miyazato M, Kangawa K.
|
| タイトル |
A guanosine 3',5'-cyclic monophosphate (cGMP) reporter system based on the G-kinase/CREB/CRE signal transduction pathway.
|
| ジャーナル |
Biochem Biophys Res Commun
|
| Abstract |
Guanylate cyclases constitute a gene family of enzymes that synthesize the second messenger guanosine 3',5'-cyclic monophosphate (cGMP) and play important roles in diverse physiological functions. Here we report a novel, simple and highly sensitive method for measurement intracellular cGMP concentrations using a cAMP-responsive element (CRE) and cGMP-dependent protein kinase (cGK). Transient transfection of the CRE reporter plasmid, encoding a luciferase reporter gene under the control of a modified promoter containing a CRE, and a cGK expression vector into HEK293 cells followed by treatment with 8-bromo-cGMP showed a dose dependent increase in luciferase activity. Moreover, HEK293 cells expressing GC-A or GC-B natriuretic peptide receptors and harboring this reporter system responded to specific ligands in a dose dependent manner. Our results indicate that this reporter gene method enables high throughput screening of receptor-type GC selective agonists in the treatment of cardiovascular diseases and homeostatic dysfunctions.
|
| 巻・号 |
407(1)
|
| ページ |
236-41
|
| 公開日 |
2011-4-1
|
| DOI |
10.1016/j.bbrc.2011.03.009
|
| PII |
S0006-291X(11)00381-0
|
| PMID |
21382339
|
| MeSH |
Cyclic AMP Response Element-Binding Protein / metabolism*
Cyclic GMP / analogs & derivatives
Cyclic GMP / analysis*
Cyclic GMP / biosynthesis
Cyclic GMP / genetics
Cyclic GMP / pharmacology
Cyclic GMP-Dependent Protein Kinase Type I
Cyclic GMP-Dependent Protein Kinases / antagonists & inhibitors
Cyclic GMP-Dependent Protein Kinases / biosynthesis*
Cyclic GMP-Dependent Protein Kinases / genetics
Drug Evaluation, Preclinical
Genes, Reporter / drug effects
HEK293 Cells
Humans
Receptors, Guanylate Cyclase-Coupled / agonists*
Response Elements
Signal Transduction
|
| IF |
2.985
|
| 引用数 |
1
|
|
WOS 分野
|
BIOPHYSICS
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
293(RCB1637) |
