| RRC ID |
22732
|
| 著者 |
Togawa S, Ishii J, Ishikura A, Tanaka T, Ogino C, Kondo A.
|
| タイトル |
Importance of asparagine residues at positions 13 and 26 on the amino-terminal domain of human somatostatin receptor subtype-5 in signalling.
|
| ジャーナル |
J Biochem
|
| Abstract |
N-linked oligosaccharides or asparagine residues are often involved in G protein-coupled receptor functions. Focusing on Asn13 and Asn26 positioned on N-linked glycosylation motifs in the amino-terminal domain of human somatostatin receptor subtype-5 (hSSTR5), we performed site-directed mutagenesis and evaluated the mutants by using yeast cells as the host strain. This is because analysing the complicated signalling in mammalian cell lines is simplified by the utilization of the monopolistic pheromone signalling pathway in yeast. Western blot analysis and confocal laser scanning microscope observation showed that Asn13 and/or Asn26 mutations had no effects on cell-surface expression of hSSTR5 in yeast. By using an engineered yeast strain of Saccharomyces cerevisiae, which induces the expression of the green fluorescent protein (GFP) reporter gene in response to the agonist-specific signal transduction, it was demonstrated that a single mutation of two asparagine residues attenuated the somatostatin-specific signalling levels, and the double mutant significantly lost the signalling ability. These results clearly show the importance of these asparagine residues in the agonist-specific signalling of hSSTR5, although it was not enough to identify the consequence of oligosaccharides.
|
| 巻・号 |
147(6)
|
| ページ |
867-73
|
| 公開日 |
2010-6-1
|
| DOI |
10.1093/jb/mvq022
|
| PII |
mvq022
|
| PMID |
20207824
|
| MeSH |
Amino Acid Substitution
Asparagine
Blotting, Western
Glycosylation
Green Fluorescent Proteins
Humans
Membrane Glycoproteins / chemistry*
Membrane Glycoproteins / metabolism
Microscopy, Confocal
Mutagenesis, Site-Directed
Receptors, G-Protein-Coupled / chemistry*
Receptors, G-Protein-Coupled / metabolism
Receptors, Somatostatin* / chemistry
Receptors, Somatostatin* / genetics
Receptors, Somatostatin* / metabolism
Saccharomyces cerevisiae
Signal Transduction / physiology*
Somatostatin / metabolism
Transfection
|
| IF |
2.476
|
| 引用数 |
20
|
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| 酵母 |
NA |
