Nadezhda V Shilova, Oxana E Galanina, Svetlana M Polyakova, Alexey Yu Nokel, Galina V Pazynina, Victoria V Golovchenko, Olga A Patova, Polina V Mikshina, Tatayana A Gorshkova, Nicolai V Bovin
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SBA, in addition to expected binding to glycans with terminal GalNAcα, also had high affinity for the GM1 ganglioside. MAH demonstrated much higher affinity to a variety of sulfated glycans compared to Neu5Acα2-3Galβ1-3GalNAcα. Contrary to the common view, LCA demonstrated the maximum binding to (GlcNAcβ1-2Manα1)<sub>2</sub>-3,6-Manβ1-4GlcNAcβ1-4GlcNAc N-glycan, while it had no interaction with corresponding Gal or Neu5Ac terminated versions. This observed polyreactivity of some lectins casts doubt on their use in accurately determining the presence of a specific glycan structure by histochemical studies. 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引用次数: 0
摘要
研究了 Jacalin、RCA 120、SBA、PHA-L、PHA-E、WGA、UEA、AAL、LTL、LEL、SNA、DSA、LCA、MAH 和 Con A 等广泛用于组织化学的凝集素的糖结合特异性。应用了基于寡糖和多糖的糖阵列。在 15 种凝集素中,只有 6 种证实了预期的特异性,而且一些凝集素的糖结合谱明显比公认的要宽。WGA、LEL 和 DSA 被称为壳寡糖特异性凝集素,但它们却意外地具有多反应性,与其他聚糖的结合亲和力与壳寡糖、ABH 抗原和低聚半乳糖胺(未取代的和硅烷基化的)相同。SBA 除了能与具有末端 GalNAcα 的聚糖结合外,还对 GM1 神经节苷脂具有很高的亲和力。与 Neu5Acα2-3Galβ1-3GalNAcα 相比,MAH 对各种硫酸化聚糖的亲和力要高得多。与通常的观点相反,LCA 与(GlcNAcβ1-2Manα1)2-3,6-Manβ1-4GlcNAcβ1-4GlcNAc N-聚糖的结合力最强,而与相应的 Gal 或 Neu5Ac 末端聚糖没有相互作用。观察到的某些凝集素的多反应性使人怀疑它们是否可用于通过组织化学研究准确确定特定聚糖结构的存在。不过,在凝集素阵列的帮助下对健康人和病人的血清进行比较,可以很容易地确定糖基化模式的差异和推测的聚糖身份,然后再用更精确的结构分析方法加以澄清。
Specificity of widely used lectins as probed with oligosaccharide and plant polysaccharide arrays.
Glycan-binding specificity was studied for Jacalin, RCA 120, SBA, PHA-L, PHA-E, WGA, UEA, AAL, LTL, LEL, SNA, DSA, LCA, MAH and Con A, lectins widely used in histochemistry. Oligosaccharide- and polysaccharide-based glycan arrays were applied. Expected specificity was confirmed for only 6 of the 15 lectins and the glycan binding profiles of some lectins were dramatically broader than generally accepted. WGA, LEL and DSA known as chitooligosaccharide-specific, were unexpectedly polyreactive, binding to other glycans with the same affinity as to chitobiose, ABH antigens and oligolactosamines (unsubstituted and sialylated). SBA, in addition to expected binding to glycans with terminal GalNAcα, also had high affinity for the GM1 ganglioside. MAH demonstrated much higher affinity to a variety of sulfated glycans compared to Neu5Acα2-3Galβ1-3GalNAcα. Contrary to the common view, LCA demonstrated the maximum binding to (GlcNAcβ1-2Manα1)2-3,6-Manβ1-4GlcNAcβ1-4GlcNAc N-glycan, while it had no interaction with corresponding Gal or Neu5Ac terminated versions. This observed polyreactivity of some lectins casts doubt on their use in accurately determining the presence of a specific glycan structure by histochemical studies. However, comparisons of sera from healthy and diseased individuals with help of a lectin array can easily establish differences in glycosylation patterns and presumptive glycan identities, which can later be clarified using more accurate methods of structural analysis.
期刊介绍:
Histochemistry and Cell Biology is devoted to the field of molecular histology and cell biology, publishing original articles dealing with the localization and identification of molecular components, metabolic activities and cell biological aspects of cells and tissues. Coverage extends to the development, application, and/or evaluation of methods and probes that can be used in the entire area of histochemistry and cell biology.