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L-fucose α1→6 specific lectin

Foreign code F120006128
File No. S2008-0721
Posted date Jan 6, 2012
Country United States of America
Application number 200913054940
Gazette No. 20110129938
Gazette No. 8461305
Date of filing Jul 15, 2009
Gazette Date Jun 2, 2011
Gazette Date Jun 11, 2013
International application number JP2009003346
International publication number WO2010010674
Date of international filing Jul 15, 2009
Date of international publication Jan 28, 2010
Priority data
  • P2008-188390 (Jul 22, 2008) JP
  • 2009WO-JP03346 (Jul 15, 2009) WO
Title L-fucose α1→6 specific lectin
Abstract (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.
Scope of claims [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.
  • Inventor, and Inventor/Applicant
  • KOBAYASHI YUKA
  • HIRABAYASHI JUN
  • TATENO HIROAKI
  • KAWAGISHI HIROKAZU
  • DOHRA HIDEO
  • J OIL MILLS
  • NATIONAL INSTITUTE OF ADVANCED INDUSTRIAL SCIENCE AND TECHNOLOGY
  • SHIZUOKA UNIVERSITY
IPC(International Patent Classification)
U.S. Cl./(Sub)
  • C07K014/37
  • C12P021/02
  • G01N033/574
  • S01N333/47A16
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