2013 Fiscal Year Final Research Report
Screening for ligands of G protein-coupled receptors using the newly developed TGFalpha shedding assay
| Project/Area Number |
24790064
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Biological pharmacy
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| Research Institution | Tohoku University |
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Principal Investigator |
INOUE Asuka 東北大学, 薬学研究科(研究院), 助手 (50525813)
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| Project Period (FY) |
2012-04-01 – 2014-03-31
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| Keywords | Gタンパク質共役型受容体 / GPCR / TGFalpha / リゾリン脂質 / リゾホスファチジルセリン / CRISPR / HEK293 / エクトドメイン切断 |
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| Research Abstract |
In this KAKENHI project, by employing the newly developed transforming growth factor-alpha (TGFalpha) shedding assay, I screened for G protein-coupled receptors (GPCRs) that respond to bioactive molecules and developed an assay system that can quantify coupling efficiency of heterotrimeric G proteins. I identified three orphan GPCRs (P2Y10, GPR174, A630033H20) as lysophoshadylserine-specific receptors, one GPCR as a lysophoshadylglucoside receptor and two GPCRs as oxidized phospholipid receptors. I generated G protein-deficient HEK293 cells using the CRISPR-Cas9 system and successfully applied them to analyze G protein coupling of GPCRs.
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[Journal Article] Overexpression of autotaxin, a lysophosphatidic acid-producing enzyme, enhances cardia bifida induced by hypo-sphingosine-1-phosphate signaling in zebrafish embryo2014
Author(s)
Nakanaga K, Hama K, Kano K, Sato T, Yukiura H, Inoue A, Saigusa D, Tokuyama H, Tomioka Y, Nishina H, Kawahara A, Aoki J
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Journal Title
Journal of Biochemistry
Volume: 155
Pages: 235-41
DOI
Peer Reviewed
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[Journal Article] TGFαshedding assay : an accurate and versatile method for detecting GPCR activation2012
Author(s)
Inoue A, Ishiguro J, Kitamura H, Arima N, Okutani M, Shuto A, Higashiyama S, Ohwada T, Arai H, Makide K, and Aoki J
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Journal Title
Nature Methods
Volume: 9
Pages: 1021-9
DOI
Peer Reviewed
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