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L-fucose α1→6 specific lectin UPDATE

外国特許コード F120006128
整理番号 S2008-0721
掲載日 2012年1月6日
出願国 アメリカ合衆国
出願番号 200913054940
公報番号 20110129938
公報番号 8461305
出願日 平成21年7月15日(2009.7.15)
公報発行日 平成23年6月2日(2011.6.2)
公報発行日 平成25年6月11日(2013.6.11)
国際出願番号 JP2009003346
国際公開番号 WO2010010674
国際出願日 平成21年7月15日(2009.7.15)
国際公開日 平成22年1月28日(2010.1.28)
優先権データ
  • 特願2008-188390 (2008.7.22) JP
  • 2009WO-JP03346 (2009.7.15) WO
発明の名称 (英語) L-fucose α1→6 specific lectin UPDATE
発明の概要(英語) (US8461305)
Disclosed is a novel lectin which can bind specifically to an L-fucose alpha1->6 sugar chain.
Also disclosed is use of the lectin.
The L-fucose alpha1->6 specific lectin of the present invention is characterized in that: (1) the lectin is extracted from a basidiomycete or an ascomycete; (2) the lectin has a molecular weight by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of 4,000 to 40,000; and (3) the lectin has an affinity to an L-fucose alpha1->6 sugar chain, the affinity being represented by an association constant of 1.0×104M-1 or more (at 25 degrees C.).
The lectin can be used for detecting an L-fucose alpha1->6 sugar chain specifically, and is effective for the purification of an L-fucose alpha1->6 sugar chain or a non L-fucose alpha1->6 sugar chain.
特許請求の範囲(英語) [claim1]
1. An L-fucose alpha 1-->6 specific lectin that: (1) is extracted from basidiomycete or ascomycete,
(2) has a molecular weight by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of 4,000 to 40,000,
(3) has an affinity to an L-fucose alpha 1-->6 sugar chain, the affinity being represented by an association constant of 1.0 * 104M-1 or more (at 25 degrees C.), and
(4) has an association constant of 1.0 * 103M-1 or less (at 25 degrees C.) for a high mannose sugar chain and/or a glycolipid not including the fucose alpha 1-->6 sugar chain.
[claim2]
2. The L-fucose alpha 1-->6 specific lectin according to claim 1, wherein the L-fucose alpha 1-->6 sugar chain has sialic acid at the nonreducing terminus.
[claim3]
3. The L-fucose alpha 1-->6 specific lectin according to claim 1, wherein the L-fucose alpha 1-->6 specific lectin (5) has an affinity to L-fucose alpha 1-->6 mono-, di-, tri-, tetra-antennary N-glycan(s), the affinity being represented by an association constant of 1.0 * 104M-1 or more (at 25 degrees C.).
[claim4]
4. The L-fucose alpha 1-->6 specific lectin according to claim 1, wherein the basidiomycete belongs to Strophariaceae, Tricholomataceae, Amanitaceae, or Polyporaceae.
[claim5]
5. The L-fucose alpha 1-->6 specific lectin according to claim 1, wherein the basidiomycete is Pholiota terrestris, Pholiota squarrosa, Pholiota adiposa, Stropharia rugosoannulata, Naematoloma sublateritium, Lepista sordida, or Amanita muscaria.
[claim6]
6. The L-fucose alpha 1-->6 specific lectin according to claim 1, wherein the L-fucose alpha 1-->6 specific lectin further (6) includes an amino acid sequence shown in SEQ ID No. 1.
[claim7]
7. An L-fucose alpha 1-->6 specific lectin that is protein or peptide consisting of an amino acid sequence shown in any of SEQ ID Nos. 2 to 6.
[claim8]
8. A method of manufacturing an L-fucose alpha 1-->6 specific lectin, comprising the steps of: subjecting an aqueous solvent extract of basidiomycete and/or ascomycete to any of following chromatography: (i) hydrophobic chromatography and reversed-phase chromatography,
(ii) affinity chromatography, and
(iii) ion-exchange chromatography and gel filtration; and
obtaining lectin having: (vi) a molecular weight by SDS-PAGE of 4,000 to 40,000,
(v) an affinity represented by an association constant to an L-fucose alpha 1-->6 sugar chain of 1.0 * 104M-1 or more, and
(vi) an affinity represented by an association constant of 1.0 * 103M-1 or less (at 25 degrees C.) for a high mannose sugar chain and/or a glycolipid which does not contain a fucose alpha 1-->6 sugar chain.
[claim9]
9. The method of manufacturing the L-fucose alpha 1-->6 specific lectin according to claim 8, wherein the basidiomycete is selected from at least one of Strophariaceae, Tricholomataceae, Amanitaceae, and Polyporaceae.
[claim10]
10. The method of manufacturing the L-fucose alpha 1-->6 specific lectin according to claim 8, wherein the basidiomycete is at least one selected from Pholiota terrestris, Pholiota squarrosa, Pholiota adiposa, Stropharia rugosoannulata, Naematoloma sublateritium, Lepista sordida, and Amanita muscaria.
[claim11]
11. The method of manufacturing the L-fucose alpha 1-->6 specific lectin according to claim 8, wherein carpophores (Fruiting bodies) of the basidiomycete and/or ascomycete are used.
[claim12]
12. A method of detecting an L-fucose alpha 1-->6 sugar chain, comprising a step of causing a sugar chain to act on the L-fucose alpha 1-->6 specific lectin according to claim 1.
[claim13]
13. The method of detecting an L-fucose alpha 1-->6 sugar chain according to claim 12, wherein the sugar chain is a tumor marker.
[claim14]
14. A method of fractionating an L-fucose alpha 1-->6 sugar chain, comprising a step of causing a sugar chain to act on the L-fucose alpha 1-->6 specific lectin according to claim 1.
[claim15]
15. The method of fractionating an L-fucose alpha 1-->6 sugar chain according to claim 14, wherein the sugar chain is bound to an antibody.
[claim16]
16. A diagnostic agent or diagnostic agent kit for detecting an L-fucose alpha 1-->6 sugar chain, wherein the diagnostic agent includes the L-fucose alpha 1-->6 specific lectin according to claim 1 as an active ingredient.
  • 発明者/出願人(英語)
  • KOBAYASHI YUKA
  • HIRABAYASHI JUN
  • TATENO HIROAKI
  • KAWAGISHI HIROKAZU
  • DOHRA HIDEO
  • J OIL MILLS
  • NATIONAL INSTITUTE OF ADVANCED INDUSTRIAL SCIENCE AND TECHNOLOGY
  • SHIZUOKA UNIVERSITY
国際特許分類(IPC)
米国特許分類/主・副
  • C07K014/37
  • C12P021/02
  • G01N033/574
  • S01N333/47A16
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