Aggregation suppression of proteins by arginine during thermal unfolding
Aggregation suppression of proteins by arginine during thermal unfolding
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DOI：
10.2174/092986606778256171
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发表时间：
2006-01-01
影响因子：
1.6
通讯作者：
Fukada, Harumi
中科院分区：
文献类型：
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作者：
Arakawa, Tsutomu;Kita, Yoshiko;Fukada, Harumi
精氨酸已被用于抑制蛋白质在重折叠和纯化过程中的聚集。我们在本文中进一步研究了精氨酸对两种商业上重要的蛋白质的聚集抑制作用，即，白细胞介素-6（IL-6）和单克隆抗体（mAb）。这些蛋白质显示出广泛的聚集在水性缓冲液中时，进行热解折叠。精氨酸抑制聚集浓度依赖性在热展开。然而，这种效果并不是精氨酸所特有的，因为相同浓度的盐酸胍（GdnHCl）也是有效的。虽然同样有效的聚集抑制在热展开过程中，精氨酸和盐酸钆不同的天然蛋白质的结构上的影响。精氨酸对天然蛋白没有明显的不利影响，而GdnHCl在室温下诱导构象变化，即，低于熔化温度。这些添加剂也影响IL-6的熔化温度;精氨酸浓度依赖性地增加IL-6的熔化温度，而GdnHCl在低浓度下增加IL-6的熔化温度，但在高浓度下降低IL-6的熔化温度。这些结果清楚地表明精氨酸通过与GdnHCl赋予的机制不同的机制抑制聚集。
Arginine has been used to suppress aggregation of proteins during refolding and purification. We have further studied in this paper the aggregation-suppressive effects of arginine on two commercially important proteins, i.e., interleukine-6 (IL-6) and a monoclonal antibody (mAb). These proteins show extensive aggregation in aqueous buffers when subjected to thermal unfolding. Arginine suppresses aggregation concentration-dependently during thermal unfolding. However, this effect was not specific to arginine, as guanidine hydrochloride (GdnHCl) at identical concentrations also was effective. While equally effective in aggregation suppression during thermal unfolding, arginine and GdnHCl differed in their effects on the structure of the native proteins. Arginine showed no apparent adverse effects on the native protein, while GdnHCl induced conformational changes at room temperature, i.e., below the melting temperature. These additives affected the melting temperature of IL-6 as well; arginine increased it concentration-dependently, while GdnHCl increased it at low concentration but decreased at higher concentration. These results clearly demonstrate that arginine suppresses aggregation via different mechanism from that conferred by GdnHCl.