Chemical synthesis of a synthetic analogue of the sialic acid-binding lectin siglec-7.

Abstract

As a basis for the development of an artificial carbohydrate-binding lectin, we chemically synthesized a domain of siglec-7, a well-characterized sialic-acid-binding lectin. The full polypeptide (127 amino acids) was constructed by sequential native chemical ligation (NCL) of five peptide segments. Because of poor cysteine availability for NCL, cysteine residues were introduced at suitable ligation sites; these cysteine residues were alkylated in order to mimic native glutamine or asparagine residues, or converted to an alanine residue by desulfurization after NCL. After folding the full-length polypeptide, the sialic-acid-binding activity of the synthetic siglec-7 was clearly demonstrated by STD NMR and ELISA experiments. We succeeded in the synthesis of siglec-7 by installing three extra cysteine residues with side-chain modifications and found that these modifications did not affect the binding activity.