...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Biotechnology >The spectroscopic analysis, inhibition and binding studies demonstrate theequivalence of Erythrina caffra trypsin inhibitor and the recombinantsubstitution variant recSerETI
【24h】

The spectroscopic analysis, inhibition and binding studies demonstrate theequivalence of Erythrina caffra trypsin inhibitor and the recombinantsubstitution variant recSerETI

机译：光谱分析，抑制和结合研究证明了刺桐尾孢胰蛋白酶抑制剂与重组取代变体recSerETI的等效性

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

A recombinant substitution mutant (recSerETI) of the Erythrina caffi a trypsin inhibitor, with the N-terminal valine residue substituted by serine, was produced in E. coli and compared to the wildtype protein (wtETI) with respect to physicochemical and functional properties. The spectral properties, including UV absorbance, fluorescence emission and circular dichroism, were indistinguishable. Furthermore, the inhibitory activities of the two proteins regarding the inhibition of trypsin, chymotrypsin, tissue plasmininogen activator (t-PA) and reteplase (BM 06.022, t-PA deletion variant comprising the kringle 2 and the protease domains, isolated from transformed E. coli cells) and the affinity of the immobilized inhibitors for reteplase were closely similar. Five repetitive cycles of guanidinium chloride (GdmCl) -induced denaturation-renaturation yield the native mutant protein with its inhibitory activity fully restored. The only difference between the wildtype and the mutant protein refers to the intrinsic stability. Comparing the pH- and GdmCl-dependent transitions, as well as the thermal denaturation, recSerETI exhibits decreased stability compared to the wildtype protein. The pH range of stability is shifted from pH 1-9.5, for wtETI, to pH 2-9, for recSerETI; similarly the GdmCl-induced denaturation is found to occur at a GdmCl half concentration of 3.7 M instead of 4.5 M; in both cases the renaturation exhibits strong hysteresis. The mid-point of the thermal unfolding transition of the mutant protein is at similar to 65 degrees C, as compared to similar to 75 degrees C for the wildtype protein.
机译：在大肠杆菌中产生了一种Erythrina caffi胰蛋白酶抑制剂的重组取代突变体（recSerETI），其N末端缬氨酸残基被丝氨酸取代，在理化和功能特性方面与野生型蛋白（wtETI）进行了比较。包括紫外吸收，荧光发射和圆二色性在内的光谱特性是无法区分的。此外，这两种蛋白在抑制胰蛋白酶，胰凝乳蛋白酶，组织纤溶酶原激活物（t-PA）和reteplase（BM 06.022，包含kringle 2和蛋白酶结构域的t-PA缺失变体）方面的抑制活性是从转化的大肠杆菌中分离出来的。大肠杆菌细胞）和固定化抑制剂对替普酶的亲和力非常相似。氯化胍（GdmCl）诱导的变性-复性的五个重复循环产生了天然的突变蛋白，其抑制活性得以完全恢复。野生型和突变蛋白之间的唯一区别是固有稳定性。比较pH和GdmCl依赖性转变以及热变性，recSerETI与野生型蛋白质相比，稳定性降低。 pH的稳定性范围从wtETI的pH 1-9.5更改为recSerETI的pH 2-9。同样，发现GdmCl诱导的变性发生在GdmCl半浓度为3.7 M而不是4.5 M时。在这两种情况下，复性均表现出很强的滞后性。与野生型蛋白质的类似于75℃相比，突变体蛋白质的热解折叠转变的中点在约65℃。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号