Premium
A novel post‐ligation thioesterification device enables peptide ligation in the N to C direction: synthetic study of human glycodelin
Author(s) -
Takenouchi Takaomi,
Katayama Hidekazu,
Nakahara Yuko,
Nakahara Yoshiaki,
Hojo Hironobu
Publication year - 2014
Publication title -
journal of peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 66
eISSN - 1099-1387
pISSN - 1075-2617
DOI - 10.1002/psc.2592
Subject(s) - native chemical ligation , peptide , chemistry , chemical ligation , cysteine , ligation , homocysteine , residue (chemistry) , combinatorial chemistry , solid phase synthesis , peptide synthesis , stereochemistry , biochemistry , enzyme , medicine
Human glycodelin consists of 162 amino acid residues and two N ‐linked glycans at Asn 28 and Asn 63 . In this study, we synthesized it by a fully convergent strategy using native chemical ligation (NCL) in N to C direction. The four peptide segments corresponding to 1–31, 32–65, 66–105 and 106–162 sequences were synthesized by 9‐fluorenylmethoxycarbonyl based solid‐phase peptide synthesis. At the C‐terminus of the second segment, N ‐ethyl‐ S ‐acetamidomethyl‐cysteine was attached as a post‐ligation thioesterification device. The N‐terminal two segments were condensed by the homocysteine‐mediated NCL at Leu‐Met site, and the product was methylated to convert homocysteine to methionine. After deprotection of acetamidomethyl group on the N ‐ethylcysteine residue, the peptide was thioesterified by N ‐alkylcysteine‐assisted method. The product was then ligated with the C‐terminal half, which was obtained by the NCL of third and fourth segments, to give the full‐length glycodelin. Copyright © 2013 European Peptide Society and John Wiley & Sons, Ltd.