Chemo-enzymatic synthesis of a bioactive peptide containing a glutamine-linked oligosaccharide and its characterization.
A bioactive peptide containing a glutamine-linked oligosaccharide was chemo-enzymatically synthesized by use of the solid-phase method of peptide synthesis and the transglycosylation activity of endo-beta-N-acetylglucosaminidase. Substance P, a neuropeptide, is an undecapeptide containing two L-glutamine residues. A substance P derivative with an N-acetyl-D-glucosamine residue attached to the fifth or sixth L-glutamine residue from the N-terminal region was chemically synthesized. A sialo complex-type oligosaccharide derived from a glycopeptide of hen egg yolk was added to the N-acetyl-D-glucosamine moiety of the substance P derivative using the transglycosylation activity of endo-beta-N-acetylglucosaminidase from Mucor hiemalis, and a substance P derivative with a sialo complex-type oligosaccharide attached to the L-glutamine residue was synthesized. This glycosylated substance P was biologically active, although the activity was rather low, and stable against peptidase digestion. The oligosaccharide moiety attached to the L-glutamine residue of the peptide was not liberated by peptide-N(4)-(N-acetyl-beta-D-glucosaminyl) asparagine amidase F.
Haneda
K
K
The Noguchi Institute, Itabashi, Tokyo, Japan. haneda@noguchi.or.jp
Inazu
T
T
Mizuno
M
M
Iguchi
R
R
Tanabe
H
H
Fujimori
K
K
Yamamoto
K
K
Kumagai
H
H
Tsumori
K
K
Munekata
E
E
D000581:Amidohydrolases; D000595:Amino Acid Sequence; D000818:Animals; D002240:Carbohydrate Sequence; D005973:Glutamine / Q000737:chemistry*; D006031:Glycosylation; D006168:Guinea Pigs; D007082:Ileum / Q000187:drug effects; D008956:Models, Chemical; D008969:Molecular Sequence Data; D009119:Muscle Contraction; D009130:Muscle, Smooth / Q000187:drug effects; D009844:Oligosaccharides / Q000737:chemistry*; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D013373:Substance P / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000494:pharmacology
D009844:Oligosaccharides; D005973:Glutamine; D013373:Substance P; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase