Precise hybrid synthesis of glycoprotein through amino acid sequence-specific introduction of oligosaccharide followed by enzymatic transglycosylation reaction

虽然它增加了我们对糖蛋白生理功能的认识，但很难获得均匀的糖蛋白。目前，利用大肠杆菌等原核细胞可以很容易地获得适当折叠的裸蛋白，并且可以通过与低聚糖和蛋白质的精确偶联来获得所需的糖蛋白。最近有报道称，低聚糖和蛋白质之间的Amadori重排以序列特异性的方式发生。基于这篇报告，我研究了添加剂对Amadori重排反应的影响以及对其反应速率的加速。最引人注目的结果是，硼酸盐离子的作用取决于糖的化学结构。例如，使用d -果糖在磷酸盐缓冲液中观察到非常大的反应速率，尽管在硼酸盐缓冲液中几乎没有观察到反应。Amadori重排的序列特异性研究正在进行中。

It is difficult to obtain homogeneous glycoproteins, although it is increasing our knowledge concerning physiological functions of glycoproteins. Nowadays, it is very easy to obtain bare proteins that fold appropriately by using prokaryote cells like Escherichia coli, and it is possible to obtain desired glycoproteins by conjugating with oligosaccharide and proteins precisely. Recently, it was reported that the Amadori rearrangement occurred between oligosaccharides and proteins in a sequence specific manner. Based on this report, I have investigated the effect of additives on the Amadori rearrangement reaction and acceleration of its reaction rate. The most striking result was that the effects of borate ion were depended upon chemical structure of sugars. For example, very large reaction rate using D-fructose was observed in phosphate buffer, although almost no reaction was observed in borate buffer. Investigation of sequence specificity of the Amadori rearrangement is under way.