Terada Takeshi, Nishikawa Makiya, Yamashita Fumiyoshi, Hashida Mitsuru
Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
Int J Pharm. 2006 Jun 19;316(1-2):117-23. doi: 10.1016/j.ijpharm.2006.02.047. Epub 2006 Apr 5.
The kinetics and specificity of the molecular interaction between proteins modified with varying numbers of mannose residues and isolated rabbit mannan-binding lectin (MBL) were characterized by using surface plasmon resonance spectroscopy (SPR). Mannosylated bovine serum albumin (Man-BSA) with different numbers of mannoses and other mannosylated derivatives of lysozyme (LZM), soybean trypsin inhibitor (STI), superoxide dismutase (SOD) and bovine gamma-immunoglobulin (IgG) were synthesized. Rabbit MBL was isolated by affinity column chromatography and immobilized on the SPR sensor chip via avidin-biotin binding. Binding of Man-BSAs to immobilized rabbit MBL increased with an increase in the number of mannose residues, primarily due to the reduction in dissociation rate. On the other hand, the association rate constant was similar for five mannosylated proteins investigated, whereas the dissociation rate constant differed markedly in spite of the same degree of mannosylation. Specific binding of mannosylated proteins to MBL may depend on the number of mannose residues and their steric configurations.
通过表面等离子体共振光谱（SPR）对不同数量甘露糖残基修饰的蛋白质与分离的兔甘露聚糖结合凝集素（MBL）之间分子相互作用的动力学和特异性进行了表征。合成了具有不同数量甘露糖的甘露糖基化牛血清白蛋白（Man-BSA）以及溶菌酶（LZM）、大豆胰蛋白酶抑制剂（STI）、超氧化物歧化酶（SOD）和牛γ-免疫球蛋白（IgG）的其他甘露糖基化衍生物。通过亲和柱色谱法分离兔MBL，并通过抗生物素蛋白-生物素结合将其固定在SPR传感器芯片上。Man-BSAs与固定化兔MBL的结合随着甘露糖残基数量的增加而增加，这主要是由于解离速率的降低。另一方面，在所研究的五种甘露糖基化蛋白质中，缔合速率常数相似，而尽管甘露糖基化程度相同，解离速率常数却有显著差异。甘露糖基化蛋白质与MBL的特异性结合可能取决于甘露糖残基的数量及其空间构型。
