Target-Specific Chemical Acylation of Lectins by Ligand-Tethered DMAP Catalysts

• Published in: Journal of the American Chemical Society Vol. 130; no. 1; pp. 245 - 251
• Main Authors: Koshi, Yoichiro, Nakata, Eiji, Miyagawa, Masayoshi, Tsukiji, Shinya, Ogawa, Tomohisa, Hamachi, Itaru
• Format: Journal Article
• Language: English
• Published: WASHINGTON American Chemical Society 09.01.2008; Amer Chemical Soc
• Subjects: Acylation; Catalysis; Chemistry; Chemistry, Multidisciplinary; Escherichia coli; Galectins - chemistry; Lectins - chemistry; Ligands; Molecular Probes; Peptide Mapping; Physical Sciences; Pyridines; Science & Technology; Staining and Labeling; Tyrosine; CONGER EEL; CARBOHYDRATE CHIPS; EVOLUTION; ARRAY; MICROARRAY; ACTIVE-SITE; ALKYLATION; CHEMISTRY; SURFACE; PROTEINS
• ISSN: 0002-7863; 1272-7863; 1520-5126
• DOI: 10.1021/ja075684q

Because sugar-binding proteins, so-called lectins, play important roles in many biological phenomena, the lectin-selective labeling should be useful for investigating biological processes involving lectins as well as providing molecular tools for analysis of saccharides and these derivatives. We describe herein a new strategy for lectin-selective labeling based on an acyl transfer reaction directed by ligand-tethered DMAP (4-dimethylaminopyridine). DMAP is an effective acyl transfer catalyst, which can activate an acyl ester for its transfer to a nucleophilic residue. To direct the acyl transfer reaction to a lectin of interest, we attached the DMAP to a saccharide ligand specific for the target lectin. It was clearly demonstrated by biochemical analyses that the target-selective labeling of Congerin II, an animal lectin having selective affinity for Lactose/LacNAc (N-acetyllactosamine), was achieved in the presence of Lac-tethered DMAPs and acyl donors containing probes such as fluorescent molecules or biotin. Conventional peptide mapping experiments using HPLC and tandem mass−mass analysis revealed that the acyl transfer reaction site-specifically occurred at Tyr 51 of Cong II. This strategy was successfully extended to other lectins by changing the ligand part of the ligand-tethered DMAP. We also demonstrated that this labeling method is applicable not only to purified lectin in test tubes, but also to crude mixtures such as E. coli lysates or homogenized animal tissue samples expressing Congerin.