Characterization of the structural determinants required for the high affinity interaction of asparagine-linked oligosaccharides with immobilized Phaseolus vulgaris leukoagglutinating and erythroagglutinating lectins.
Characterization of the structural determinants required for the high affinity interaction of asparagine-linked oligosaccharides with immobilized Phaseolus vulgaris leukoagglutinating and erythroagglutinating lectins.
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DOI：
10.1016/s0021-9258(18)33746-3
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发表时间：
1982-10
期刊：
影响因子：
--
通讯作者：
R. Cummings;S. Kornfeld
中科院分区：
文献类型：
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作者：
R. Cummings;S. Kornfeld
用天冬氨酸连接的低聚糖凝集素-琼脂糖亲和层析法研究了四季豆白血球凝集素(L-PHA)和红细胞凝集素(E-PHA)凝集素的糖结合特性。只有含有2个外层半乳糖残基和N-乙酰氨基葡萄糖残基的双天线糖肽才能与E-PHA-琼脂糖高亲和力结合，N-乙酰氨基葡萄糖残基与核心中的β-甘露糖残基相连。本种在L-PHA-琼脂糖凝胶上不迟钝。相反，含有外层半乳糖残基和C-2和C-6位取代的α-甘露糖残基的三元和四元糖肽在L-PHA-琼脂糖凝胶上被特异性地延缓。含有外层半乳糖残基和C-2和C-4位取代的α-甘露糖残基的三天线糖肽在L-PHA-琼脂糖凝胶上不被阻挡。此外，外层唾液酸残基或核心岩藻糖残基的存在不会影响复合糖肽在这些凝集素-琼脂糖偶联物上的行为。E-PHA和L-PHA能够区分具有不同分支模式的天冬氨酸连接的寡糖，这一能力可用于这些糖肽的分离(见下文)。
The carbohydrate binding specificities of the leukoagglutinating phytohemagglutinin (L-PHA) and erythroagglutinating phytohemagglutinin (E-PHA) lectins of the red kidney bean, Phaseolus vulgaris, have been investigated by lectin-agarose affinity chromatography of Asn-linked oligosaccharides. High affinity binding to E-PHA-agarose occurs only with biantennary glycopeptides containing 2 outer galactose residues and a residue of N-acetylglucosamine linked beta 1,4 to the beta-linked mannose residue in the core. This species is not retarded on L-PHA-agarose. In contrast, tri- and tetraanternnary glycopeptides containing outer galactose residues and an alpha-linked mannose residue substituted at positions C-2 and C-6 are specifically retarded on L-PHA-agarose. Triantennary glycopeptides containing outer galactose residues and an alpha-linked mannose residue substituted at positions C-2 and C-4 are not retarded on L-PHA-agarose. Additionally, the presence of outer sialic acid residues or a core fucose residue does not influence the behavior of complex glycopeptides on either of these lectin-agarose conjugates. This ability of E-PHA and L-PHA to discriminate between Asn-linked oligosaccharides with various branching patterns can be utilized in the fractionation of these glycopeptides (see paper following).