SYNTHESIS OF 1-N-GLYCYL BETA-OLIGOSACCHARIDE DERIVATIVES - REACTIVITY OF LENS-CULINARIS LECTIN WITH A FLUORESCENT LABELED STREPTAVIDIN PSEUDOGLYCOPROTEIN AND IMMOBILIZED NEOGLYCOLIPID
SYNTHESIS OF 1-N-GLYCYL BETA-OLIGOSACCHARIDE DERIVATIVES - REACTIVITY OF LENS-CULINARIS LECTIN WITH A FLUORESCENT LABELED STREPTAVIDIN PSEUDOGLYCOPROTEIN AND IMMOBILIZED NEOGLYCOLIPID
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DOI：
10.1021/bi00159a013
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发表时间：
1992-11-10
期刊：
影响因子：
2.9
通讯作者：
DWEK, RA
中科院分区：
文献类型：
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作者：
MANGER, ID;WONG, SYC;DWEK, RA
小豆凝集素(Lens culinaris，Lentil)与复合型N-连接低聚糖的还原末端含有6-O-连接岩藻糖的糖肽具有结合特异性。因此，小扁豆凝集素提供了一个很好的碳水化合物结合蛋白的例子，其中高亲和力的相互作用依赖于低聚糖核心结构的完整性。本文报道了Galbeta4GlcNAcbeta2Manalpha6(Galbeta4GlcNAcbeta2Manalpha3)Manbeta4GlcNAcbeta4(Fucalpha6)GlcNAc的1-N-甘氨酰β-衍生物(GAL-2F)的合成及其生物素化和棕榈酰化反应。当生物素衍生物与链霉亲和素-异硫氰酸酯(FITC)偶联物结合时，能够与刀豆蛋白A(ConA)和小扁豆凝集素亲和层析柱结合。相反，合成Gal-2F的甘氨酸衍生物的生物素衍生物并随后与链霉亲和素-FITC结合后，可与ConA亲和柱反应，但不与小扁豆凝集素亲和柱反应。扁豆凝集素还结合到含有吸附的棕榈酰基-1-N-甘氨酰-β-衍生物的塑料微滴定板上。当使用同源糖胺新糖脂时，没有发生结合。这些结果表明，低聚糖的1-N-甘氨酰-β-衍生物在合成新型糖共轭探针方面可能具有广泛的用途。
The lectin from Lens culinaris (lentil) has a binding specificity for glycopeptides bearing 6-O-linked fucose on the reducing terminus on complex-type N-linked oligosaccharides. Lentil lectin therefore provides an excellent example of a carbohydrate binding protein in which high-affinity interactions are dependent on the integrity of the oligosaccharide core structure. We report here the synthesis of the 1-N-glycyl beta-derivative of Galbeta4GlcNAcbeta2Manalpha6(Galbeta4GlcNAcbeta2Manalpha3)Manbeta4GlcNAcbeta4(Fucalpha6)GlcNAc (Gal-2F) and its subsequent biotinylation and palmitoylation. The biotin derivative when bound to a streptavidin-fluorescein isothiocyanate (FITC) conjugate was able to bind to both concanavalin A (ConA) and lentil lectin affinity columns. In contrast, synthesis of the biotin derivative of the glycamine derivative of Gal-2F and subsequent binding to streptavidin-FITC afforded reactivity to a ConA affinity column but not to a lentil lectin affinity column. Lentil lectin also bound to plastic microtiter plates containing the adsorbed palmitoyl-1-N-glycyl beta-derivative. No binding occurred when the homologous glycamine neoglycolipid was used. These results suggest the 1-N-glycyl beta-derivative of oligosaccharides may have general utility as an intermediate in the synthesis of novel glycoconjugate probes.