Shiraki Kentaro, Kudou Motonori, Sakamoto Ryusuke, Yanagihara Itaru, Takagi Masahiro
School of Materials Science, Japan Advanced Institute of Science and Technology, Tatsunokuchi, Ishikawa 923-1292, Japan.
Biotechnol Prog. 2005 Mar-Apr;21(2):640-3. doi: 10.1021/bp049769w.
Small potent inhibitors of aggregation are eagerly demanded for preventing the inactivation of proteins. This paper shows that amino acid esters (AAEs) prevent heat-induced aggregation and inactivation of hen egg lysozyme. Lysozyme was completely inactivated (<1% original activity) during heat treatment at 98 degrees C for 30 min in a solution containing 0.2 mg/mL lysozyme in 50 mM Na-phosphate buffer (pH 6.5). The residual activities only slightly increased (<5%) in the presence of 100 mM commonly used additives such as arginine, guanidine, urea, and sugars. However, in the presence of 100 mM AAEs, the residual activities were >60% and no aggregates were observed during the heat treatment at 98 degrees C for 30 min. This fact provides new information on the scaffold for designing additives to prevent heat-induced aggregation.
为防止蛋白质失活,人们迫切需要高效的小分子聚集抑制剂。本文表明,氨基酸酯(AAEs)可防止热诱导的鸡蛋清溶菌酶聚集和失活。在含有0.2 mg/mL溶菌酶的50 mM磷酸钠缓冲液(pH 6.5)中,于98℃热处理30分钟后,溶菌酶完全失活(活性低于原始活性的1%)。在存在100 mM常用添加剂(如精氨酸、胍、尿素和糖)的情况下,残余活性仅略有增加(<5%)。然而,在存在100 mM AAEs的情况下,残余活性>60%,并且在98℃热处理30分钟期间未观察到聚集体。这一事实为设计防止热诱导聚集的添加剂支架提供了新信息。