TOP > 外国特許検索 > Probe for detecting and quantifying lipid second messenger and method of detecting and quantifying lipid second messenger using the same

Probe for detecting and quantifying lipid second messenger and method of detecting and quantifying lipid second messenger using the same 実績あり

外国特許コード F110005209
整理番号 A161-08US
掲載日 2011年8月26日
出願国 アメリカ合衆国
出願番号 55419404
公報番号 20060265764
公報番号 7507555
出願日 平成16年3月15日(2004.3.15)
公報発行日 平成18年11月23日(2006.11.23)
公報発行日 平成21年3月24日(2009.3.24)
国際出願番号 JP2004003433
国際公開番号 WO2004095024
国際出願日 平成16年3月15日(2004.3.15)
国際公開日 平成16年11月4日(2004.11.4)
優先権データ
  • 特願2003-120253 (2003.4.24) JP
  • 2004WO-JP03433 (2004.3.15) WO
発明の名称 (英語) Probe for detecting and quantifying lipid second messenger and method of detecting and quantifying lipid second messenger using the same 実績あり
発明の概要(英語) (US7507555)
The present invention provides probes for detection and quantification of a lipid second messenger, which comprises: a polypeptide specifically bound to the lipid second messenger; two chromophores respectively having different fluorescence wavelengths, wherein each of the chromophores is linked to each end of the polypeptide through a rigid linker sequence; and a membrane localization sequence linked to one of the chromophores through a rigid linker sequence.
According to the present invention, it is now possible to quantitatively detect when and in which site of a living cell the lipid second messengers are produced.
特許請求の範囲(英語) [claim1]
1. A probe for detection and quantification of a lipid second messenger, which comprises: a polypeptide which can specifically bind the lipid second messenger,a first chromophore linked to one end of the polypeptide through a rigid linker sequence;
a second chromophore linked to another end of the polypeptide through a second rigid linker sequence, wherein the second chromophore has a different fluorescence wavelength from the first chromophore, and the second linker sequence is rigid except for a single flexible site acting as a hinge;
and
a membrane localization sequence linked to the second chromophore through a third rigid linker sequence, wherein when the polypeptide is bound to the lipid second messenger, the first and second chromophores are capable of Fluorescence Resonance Energy Transfer (FRET).
[claim2]
2. The probe for detection and quantification of a lipid second messenger of claim 1, wherein the polypeptide which can specifically bind the lipid second messenger is a lipid second messenger-binding protein.
[claim3]
3. The probe for detection and quantification of a lipid second messenger of claim 2, wherein the lipid second messenger-binding protein is a pleckstrin homology domain from General Receptor for Phosphoinositides-1 (GRP 1).
[claim4]
4. The probe for detection and quantification of a lipid second messenger of claim 1, wherein the chromophores are a cyan fluorescent protein linked to N-terminal end of the polypeptide and a yellow fluorescent protein linked to C-terminal end of the polypeptide.
[claim5]
5. The probe for detection and quantification of a lipid second messenger of claim 1, wherein the first, second and third linker sequences comprise a rigid alpha -helix linker consisting of repeated sequences of SEQ ID NO: 1.
[claim6]
6. The probe for detection and quantification of a lipid second messenger of claim 1, wherein the single flexible site of the second linker sequence is a single di-glycine motif.
[claim7]
7. The probe for detection and quantification of a lipid second messenger of claim 1, wherein the membrane localization sequence is a lipidized sequence or a transmembrane sequence.
[claim8]
8. A method for detecting and quantifying a lipid second messenger, comprising: contacting the probe with the lipid second messenger of claim 1;
and measuring changes in fluorescence spectra.
  • 発明者/出願人(英語)
  • UMEZAWA YOSHIO
  • SATO MORITOSHI
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
国際特許分類(IPC)
米国特許分類/主・副
  • 435/69.1
  • 435/15
  • 435/69.7
  • 435/194
  • 436/94
  • 436/501
参考情報 (研究プロジェクト等) CREST Endocrine Disrupters AREA
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