比較兩種甘露糖／葡萄糖專一性之刀豆凝集素及碗豆凝集素的結合性質

Comparison of Binding Profile of Two Man/Glc Specific Lectins-Concanavalin A (Con A) and Pisum Sativum Lectin (PSA)

蔡明松(Ming-Sung Tsai) ； 吳千槿(Chien-Ching Wu)

《化學》 68卷3期 (2010/09) Pp. 235-246

https://doi.org/10.6623/chem.2010022

摘要

本研究主要利用酵素連結凝集素分析法及抑制分析法研究刀豆凝集素和碗豆凝集素與不同N-鍵結醣蛋白、O-鍵結醣蛋白及多醣體的結合，在抑制分析法中用寡糖、單糖及其衍生物作為抑制劑。實驗結果顯示刀豆凝集素與碗豆凝集素主要偏愛N-鍵結醣蛋白，但所測試的O-鍵結醣蛋白中刀豆凝集素亦能與包含II構造之O-鍵結醣蛋白如酸性水解與不水解的豬胃黏液醣蛋白、去唾液酸之燕窩醣蛋白及肺炎鏈球菌莢膜多醣體，而丁二酸醯化刀豆凝集素卻與O-鍵結醣蛋白都沒有活性，而碗豆凝集素僅對去唾液酸之燕窩醣蛋白有反應。另外當GlcNAc鍵結到甘露糖三元體會降低對碗豆凝集素的抑制力，但反而提高對刀豆凝集素的抑制能力，顯示刀豆凝集素有一個能結合GlcNAc的次結合位置。最後比較刀豆凝集素及丁二酸醯化刀豆凝集素結合能力顯示多價結合作用對具四個結合位置的刀豆凝集素與碳水化合物的結合貢獻比僅有二個結合位置的丁二酸醯化刀豆凝集素大。

關鍵字

刀豆凝集素 ； 碗豆凝集素 ； 丁二酸醯化刀豆凝集素

並列摘要

Concanavalin A, Pisum sativum lectin are specific for Man/Glc, but differ in the effect of glycan polyvalency on both affinity and specificity, and their binding relationship towards mammalian disaccharide carbohydrate structural units have not been examined. In this study, all recognition factors at the combing sites Con A and PSA were analyzed by enzyme linked lectinosorbent and inhibition assays with a panel of natural polyvalent glycans and an array of mono-, di-, oligosaccharides and mammalian carbohydrate structural units. Based on the results of this study, it can be concluded that: (a) very high affinity exit between Con A, PSA and oligomannosyl structural units of N-glycans; (b) The reactivities for mannose/glucose polysaccharide (Mannan/ glycogen) were good ligand for Con A, while PSA and succinyl-Con A showed were weak or inactive; (c) Con A and PSA were weak or inactive with O-glycans, but blood group A+Hcontaining glycoproteins and their mild-acid hydrolysed products from hog stomach mucin would well react with Con A, while PSA and succinyl-Con A were inactive; (d) Comparisons of binding of Con A, PSA and succinyl-Con A to various gps and polysaccharides by ELLSA showed that carbohydrate-binding affinity of Con A was better than succinyl-Con A and PSA, respectively; (e) the recognition specificity of Con A and PSA for oligosaccharides can be defined in decreasing order as follows: Con A-α1-3, α1-6 Mannopetaose, biantennary N-linked core pentasaccharide ＞ α1-3, α1-6 Mannotriose ＞ Threhalose ＞ Manα1-3Man ＞ Manα1-2Man ＞ Manα1-6Man ＞ Raffinose ＞ Melibiose ＞ GlcNAcβ1-2Man ＞ Tri-II ＞ Maltotriose ＞ Maltose ＞ Strachyose, E, T, S, B, F, Tn, I, P, A, II and L were inactive. However, PSA-α1-3, α1-6 Mannopetaose ＞ Manα1- 2Man ＞ α1-3, α1-6 Mannotriose ＞ biantennary N-linked core pentasaccharide, Manα1-3Man ＞ Manα1-4Man ＞ Threhalose ＞ Manα1-6Man ＞ Maltose ＞ GlcNAcβ1-2Man ＞ Maltotriose ＞ Raffinose ＞ Melibiose ＞ Tri-II, E, T, S, B, F, Tn, I, P, A, II and L were inactive.

並列關鍵字

Concanavalin A ； Pisum sativum lectin ； Succinyl-Con A ； ELLSA