Reaction #979164

ord-10c9037bc0314be9952ae4dbb5aff82b

Reaction equation

CC(=O)NC1C(OC2C(O)C(CO)OC(OC(C(O)CO)C(OC3OC(C)C(O)C(O)C3O)C(O)C=O)C2O)OC(CO)C(O)C1OC1OC(CO)C(O)C(O)C1O
LNFP-V
CC(=O)N[C@H]1[C@H](O[C@H]2[C@@H](O)[C@@H](CO)O[C@@H](O[C@H]3[C@H](O)[C@@H](O)C(O)O[C@@H]3CO)[C@@H]2O)O[C@H](CO)[C@@H](O[C@@H]2O[C@H](CO)[C@H](O)[C@H](O)[C@H]2O)[C@@H]1O
LNnT
CC(=O)N[C@H]1[C@H](O[C@H]2[C@@H](O)[C@@H](CO)O[C@@H](O[C@H]3[C@H](O[C@@H]4O[C@@H](C)[C@@H](O)[C@@H](O)[C@@H]4O)[C@@H](O)C(O)O[C@@H]3CO)[C@@H]2O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O
Galβ1-3GlcNAcβ1-3Galβ1-4(Fucα1-3)Glc
CC(=O)N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O
N-acetylglucosamine
CC(=O)N[C@H]1[C@H](O[C@H]2[C@@H](O)[C@@H](CO)O[C@@H](O[C@H]3[C@H](O)[C@@H](NC(C)=O)[C@H](O[C@H]4[C@@H](O)[C@@H](CO)O[C@@H](O[C@H]5[C@H](O)[C@@H](O)C(O)O[C@@H]5CO)[C@@H]4O)O[C@@H]3CO)[C@@H]2O)O[C@H](CO)[C@@H](O[C@@H]2O[C@H](CO)[C@H](O)[C@H](O)[C@H]2O)[C@@H]1O
Galβ1-4GlcNAcβ1-3Galβ1-4GlcNAcβ1-3Galβ1-4Glc
→
CC(=O)N[C@H]1[C@H](O[C@H]2[C@@H](O)[C@@H](CO)O[C@@H](O[C@H]3[C@H](O)[C@@H](CO)O[C@@H](O[C@H]4[C@@H](O)[C@@H](CO)O[C@@H](O[C@H]5[C@H](O[C@@H]6O[C@@H](C)[C@@H](O)[C@@H](O)[C@@H]6O)[C@@H](O)C(O)O[C@@H]5CO)[C@@H]4O)[C@@H]3NC(C)=O)[C@@H]2O)O[C@H](CO)[C@@H](O[C@@H]2O[C@H](CO)[C@H](O)[C@H](O)[C@H]2O)[C@@H]1O
Galβ1-4GlcNAcβ1-3Galβ1-3GlcNAcβ1-3Galβ1-4(Fucα1-3)Glc

Reactants

—
polypeptide
CC(=O)NC1C(OC2C(O)C(CO)OC(OC(C(O)CO)C(OC3OC(C)C(O)C(O)C3O)C(O)C=O)C2O)OC(CO)C(O)C1OC1OC(CO)C(O)C(O)C1O
LNFP-V
CC(=O)N[C@H]1[C@H](O[C@H]2[C@@H](O)[C@@H](CO)O[C@@H](O[C@H]3[C@H](O)[C@@H](O)C(O)O[C@@H]3CO)[C@@H]2O)O[C@H](CO)[C@@H](O[C@@H]2O[C@H](CO)[C@H](O)[C@H](O)[C@H]2O)[C@@H]1O
LNnT
CC(=O)N[C@H]1[C@H](O[C@H]2[C@@H](O)[C@@H](CO)O[C@@H](O[C@H]3[C@H](O[C@@H]4O[C@@H](C)[C@@H](O)[C@@H](O)[C@@H]4O)[C@@H](O)C(O)O[C@@H]3CO)[C@@H]2O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O
Galβ1-3GlcNAcβ1-3Galβ1-4(Fucα1-3)Glc
CC(=O)N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O
N-acetylglucosamine
CC(=O)N[C@H]1[C@H](O[C@H]2[C@@H](O)[C@@H](CO)O[C@@H](O[C@H]3[C@H](O)[C@@H](NC(C)=O)[C@H](O[C@H]4[C@@H](O)[C@@H](CO)O[C@@H](O[C@H]5[C@H](O)[C@@H](O)C(O)O[C@@H]5CO)[C@@H]4O)O[C@@H]3CO)[C@@H]2O)O[C@H](CO)[C@@H](O[C@@H]2O[C@H](CO)[C@H](O)[C@H](O)[C@H]2O)[C@@H]1O
Galβ1-4GlcNAcβ1-3Galβ1-4GlcNAcβ1-3Galβ1-4Glc
—
peptide

Reagents

None

Conditions

Detailed conditions
See reaction.notes.procedure_details.

Workup

  1. 1
    Otherwas synthesized
  2. 2
    Otherproduced as a secreted form in Example 10
  3. 3
    Otherpurified from bovine milk
  4. 4
    OtherIn the same manner, Galβ1-4GlcNAcβ1-3Galβ1-3GlcNAcβ1-3Galβ1-4Glc was synthesized
  5. 5
    OtherThe specific reaction

Procedure

A sugar chain in which N-acetylglucosamine was added to the non-reducing end of the sugar chain (Galβ1-4GlcNAcβ1-3Galβ1-4GlcNAcβ1-3Galβ1-4Glc) was synthesized by allowing the FLAG peptide-fused G4 polypeptide produced as a secreted form in Example 10 to react simultaneously with LNnT and β1,4-galactosyltransferase (manufactured by SIGMA) purified from bovine milk. In the same manner, Galβ1-4GlcNAcβ1-3Galβ1-3GlcNAcβ1-3Galβ1-4Glc was synthesized using LNT as a substrate. Also, Galβ1-4GlcNAcβ1-3Galβ1-3GlcNAcβ1-3Galβ1-4(Fucα1-3)Glc was synthesized by using LNFP-V [(Galβ1-3GlcNAcβ1-3Galβ1-4(Fucα1-3)Glc) as a substrate. The specific reaction was carried out as follows.

Source

DOI: 10.6084/m9.figshare.5104873.v1Patent: US07005279B1uspto-grants-2006_02