Chemical synthesis of a glycoprotein having an intact human complex-type sialyloligosaccharide under the Boc and Fmoc synthetic strategies.
The chemical synthesis of complex glycoproteins is an ongoing challenge in protein chemistry. We have examined the synthesis of a single glycoform of monocyte chemotactic protein-3 (MCP-3), a CC-chemokine that consists of 76 amino acids and one N-glycosylation site. A three-segment native chemical ligation strategy was employed using unprotected peptides and glycopeptide. Importantly, the synthesis required the development of methods for the generation of sialylglycopeptide-alphathioesters. For the sialylglycopeptide-alphathioester segment, we examined and successfully implemented approaches using Fmoc-SPPS and Boc-SPPS. To avoid use of hydrogen fluoride, the Boc approach utilized minimal side chain protection and direct thiolysis of the resin bound peptide. Using these strategies, we successfully synthesized a glycoprotein having an intact and homogeneous complex-type sialyloligosaccharide.
Yamamoto
Naoki
N
International Graduate School of Arts and Sciences, Yokohama City University, 22-2, Seto, Kanazawa-ku, Yokohama, 236-0027 Japan.
Tanabe
Yasutaka
Y
Okamoto
Ryo
R
Dawson
Philip E
PE
Kajihara
Yasuhiro
Y
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry*; D054410:Chemokine CCL7 / Q000138:chemical synthesis*; D005449:Fluorenes / Q000737:chemistry*; D005563:Formic Acid Esters / Q000737:chemistry*; D006023:Glycoproteins / Q000138:chemical synthesis*; D006801:Humans; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry*
D000596:Amino Acids; D054410:Chemokine CCL7; D005449:Fluorenes; D005563:Formic Acid Esters; D006023:Glycoproteins; C016456:N(alpha)-fluorenylmethyloxycarbonylamino acids; D009844:Oligosaccharides; C016735:sialooligosaccharides; C000345:t-butyloxycarbonyl group