TY - JOUR
T1 - Lectinochemical studies on the affinity of Anguilla anguilla agglutinin for mammalian glycotopes
AU - Wu, Albert M.
AU - Wu, June H.
AU - Singh, Tanuja
AU - Liu, Jia Hau
AU - Herp, Anthony
PY - 2004/7/16
Y1 - 2004/7/16
N2 - Anguilla anguilla agglutinin (AAA) is a fucose-specific lectin found in the serum of the fresh water eel. It is suggested to be associated with innate immunity by recognizing disease-associated cell surface glycans, and has been widely used as a reagent in hematology and glycobiology. In order to gain a better understanding of AAA for further applications, it is necessary to elucidate its binding profile with mammalian glycotopes. We, therefore, analyzed the detailed carbohydrate specificity of AAA by enzyme-linked lectinosorbent assay (ELLSA) with our extended glycan/ligand collection and lectin-glycan inhibition assay. Among the glycans tested, AAA reacted well with nearly all human blood group Ah (GalNAcα1→3[LFucα1→2]Gal), Bh (Galα1→3[LFucα1→2]Gal), H LFucα1→2Gal) and Leb (Fucα1→2Galβ1→ 3[Fucα1→4]GlcNAc) active glycoproteins (gps), but not with blood group Lea (Galβ1→3[Fucα1→4]GlcNAc) substances, suggesting that residues and optimal density of α1-2 linked LFuc to Gal at the non-reducing end of glycoprotein ligands are essential for lectin-carbohydrate interactions. Blood group precursors, Galβ1-3GalNAc (T), GalNAcα1-Ser/Thr (Tn) containing glycoproteins and N-linked plasma gps, gave only negligible affinity. Among the mammalian glycotopes tested, Ah, Bh and H determinants were the best, being about 5 to 6.7 times more active than LFuc, but were weaker than p-nitrophenylαFuc indicating that hydrophobic environment surrounding the LFuc moiety enhance the reactivity. The hierarchy of potency of oligo- and monosaccharides can be ranked as follows: p-nitrophenyl-αFuc > Ah, Bh and H > LFuc > LFucα1→2Galβ1→4Glc (2′-FL) and Galβ1→4[LFucα1→3]Glc (3′-FL), while LNDFH I (Le b hexa-), Lea, Lex (Galβ1→ 4[Fucα1→3]GlcNAc), and LDFT (gluco-analogue of Ley) were inactive. From the present observations, it can be concluded that the combining site of AAA should be a small cavity-type capable of recognizing mainly H/crypto H and of binding to specific polyvalent ABH and Leb glycotopes.
AB - Anguilla anguilla agglutinin (AAA) is a fucose-specific lectin found in the serum of the fresh water eel. It is suggested to be associated with innate immunity by recognizing disease-associated cell surface glycans, and has been widely used as a reagent in hematology and glycobiology. In order to gain a better understanding of AAA for further applications, it is necessary to elucidate its binding profile with mammalian glycotopes. We, therefore, analyzed the detailed carbohydrate specificity of AAA by enzyme-linked lectinosorbent assay (ELLSA) with our extended glycan/ligand collection and lectin-glycan inhibition assay. Among the glycans tested, AAA reacted well with nearly all human blood group Ah (GalNAcα1→3[LFucα1→2]Gal), Bh (Galα1→3[LFucα1→2]Gal), H LFucα1→2Gal) and Leb (Fucα1→2Galβ1→ 3[Fucα1→4]GlcNAc) active glycoproteins (gps), but not with blood group Lea (Galβ1→3[Fucα1→4]GlcNAc) substances, suggesting that residues and optimal density of α1-2 linked LFuc to Gal at the non-reducing end of glycoprotein ligands are essential for lectin-carbohydrate interactions. Blood group precursors, Galβ1-3GalNAc (T), GalNAcα1-Ser/Thr (Tn) containing glycoproteins and N-linked plasma gps, gave only negligible affinity. Among the mammalian glycotopes tested, Ah, Bh and H determinants were the best, being about 5 to 6.7 times more active than LFuc, but were weaker than p-nitrophenylαFuc indicating that hydrophobic environment surrounding the LFuc moiety enhance the reactivity. The hierarchy of potency of oligo- and monosaccharides can be ranked as follows: p-nitrophenyl-αFuc > Ah, Bh and H > LFuc > LFucα1→2Galβ1→4Glc (2′-FL) and Galβ1→4[LFucα1→3]Glc (3′-FL), while LNDFH I (Le b hexa-), Lea, Lex (Galβ1→ 4[Fucα1→3]GlcNAc), and LDFT (gluco-analogue of Ley) were inactive. From the present observations, it can be concluded that the combining site of AAA should be a small cavity-type capable of recognizing mainly H/crypto H and of binding to specific polyvalent ABH and Leb glycotopes.
KW - Anguilla anguilla
KW - Carbohydrate specificity
KW - Combining site
KW - Glycoproteins
KW - Lectins
UR - http://www.scopus.com/inward/record.url?scp=2942648240&partnerID=8YFLogxK
U2 - 10.1016/j.lfs.2004.02.016
DO - 10.1016/j.lfs.2004.02.016
M3 - 文章
C2 - 15207656
AN - SCOPUS:2942648240
SN - 0024-3205
VL - 75
SP - 1085
EP - 1103
JO - Life Sciences
JF - Life Sciences
IS - 9
ER -
