晶体学和溶液散射分析揭示破伤风神经毒素的结构柔韧性

IF 3.5 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Chun-ming Zhang , Yoshihiro Imoto , Takaaki Hikima , Tsuyoshi Inoue
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引用次数: 4

摘要

尽管破伤风神经毒素(TeNT)通过其受体结合域(HC)和易位域(HN)将其蛋白酶结构域(LC)穿过突触囊泡腔进入细胞质,但协调这种膜易位的分子机制尚不清楚。本文报道了全长还原型TeNT (rTeNT, 2.3 Å)、TeNT分离型HN (TeNT/iHN, 2.3 Å)、TeNT分离型HC (TeNT/iHC, 1.5 Å)的高分辨率晶体结构,以及TeNT/iHN和无带型TeNT/iHN的溶液结构(TeNT/blHN)。通过对rTeNT和非还原-TeNT (nrTeNT)的结构比较,证明了ntent在HN和LC中存在显著的结构域旋转。连接HN和HC的连接环对于TeNT的自畴旋转是必不可少的。tent特异性的C869-C1093二硫键对氧化还原环境敏感,其破坏提供了连接环的灵活性,这使得rTeNT的结构域排列与nrTeNT不同。此外,通过TeNT/iHC晶体结构分析证实了C869在连接环中的迁移率和C1093对氧化还原条件的敏感性。另一方面,通过晶体学和溶液散射分析研究了HN的结构柔韧性。结果发现,位于易位区之后的区域(残基698-769)在TeNT/iHN中发生了显著变化。此外,在酸性ph下,所谓的带区具有在TeNT/iHN上半部分附近摆动的高倾向。它提供了溶液中带动力学的第一个概述。这些新获得的结构信息揭示了TeNT的跨膜机制。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Structural flexibility of the tetanus neurotoxin revealed by crystallographic and solution scattering analyses

Structural flexibility of the tetanus neurotoxin revealed by crystallographic and solution scattering analyses

Although the tetanus neurotoxin (TeNT) delivers its protease domain (LC) across the synaptic vesicle lumen into the cytosol via its receptor binding domain (HC) and translocation domain (HN), the molecular mechanism coordinating this membrane translocation remains unresolved. Here, we report the high-resolution crystal structures of full-length reduced TeNT (rTeNT, 2.3 Å), TeNT isolated HN (TeNT/iHN, 2.3 Å), TeNT isolated HC (TeNT/iHC, 1.5 Å), together with the solution structures of TeNT/iHN and beltless TeNT/iHN (TeNT/blHN). TeNT undergoes significant domains rotation of the HN and LC were demonstrated by structural comparison of rTeNT and non-reduced-TeNT (nrTeNT). A linker loop connects the HN and HC is essential for the self-domain rotation of TeNT. The TeNT-specific C869-C1093 disulfide bond is sensitive to the redox environment and its disruption provides linker loop flexibility, which enables domain arrangement of rTeNT distinct from that of nrTeNT. Furthermore, the mobility of C869 in the linker loop and the sensitivity to redox condition of C1093 were confirmed by crystal structure analysis of TeNT/iHC. On the other hand, the structural flexibility of HN was investigated by crystallographic and solution scattering analyses. It was found that the region (residues 698–769), which follows the translocation region had remarkable change in TeNT/iHN. Besides, the so-called belt region has a high propensity to swing around the upper half of TeNT/iHN at acidic pH. It provides the first overview of the dynamics of the Belt in solution. These newly obtained structural information that shed light on the transmembrane mechanism of TeNT.

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来源期刊
Journal of Structural Biology: X
Journal of Structural Biology: X Biochemistry, Genetics and Molecular Biology-Structural Biology
CiteScore
6.50
自引率
0.00%
发文量
20
审稿时长
62 days
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