Fast preparation of an N-acetylglucosaminylated peptide segment for the chemoenzymatic synthesis of a glycoprotein.
A novel GlcNAc-Asn unit carrying trifluoroacetic acid (TFA)-sensitive O-protecting groups was prepared. The unit was used for the solid-phase peptide synthesis (SPPS) of the N-acetylglucosaminylated emmprin (35-69) thioester via one-step deprotection by TFA combined with the N-alkylcysteine thioesterification method. This segment was used for the synthesis of the first Ig domain (22-104) of emmprin carrying GlcNAc by one-pot ligation with other segments using the thioester method. Finally, the sugar chain was elongated by transglycosylation using glycosynthase to give the Ig domain carrying the disialo- and asialo-complex-type sugar chain.
Department of Applied Biochemistry, Tokai University, Hiratsuka, Kanagawa 259-1292, Japan.
Yuya
Y
Asahina
Mika
M
Kanda
Akemi
A
Suzuki
Hidekazu
H
Katayama
Yoshiaki
Y
Nakahara
Hironobu
H
Hojo
D005944:Glucosamine / Q000031:analogs & derivatives / Q000138:chemical synthesis* / Q000737:chemistry*; D006023:Glycoproteins / Q000096:biosynthesis* / Q000737:chemistry; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000378:metabolism*; D006031:Glycosylation; D015394:Molecular Structure; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry*
D006023:Glycoproteins; D010455:Peptides; D006026:Glycoside Hydrolases; D005944:Glucosamine