Investigation of the Effects of Primary Structure Modifications within the RRE Motif on the Conformation of Synthetic Bovine Herpesvirus 1‐Encoded UL49.5 Protein Fragments,Chemistry & Biodiversity

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Investigation of the Effects of Primary Structure Modifications within the RRE Motif on the Conformation of Synthetic Bovine Herpesvirus 1‐Encoded UL49.5 Protein Fragments
Chemistry & Biodiversity ( IF 2.3 ) Pub Date : 2021-01-11 , DOI: 10.1002/cbdv.202000883
Natalia Karska _{1,

2} , Małgorzata Graul ₂ , Emilia Sikorska ₁ , Magdalena J Ślusarz ₁ , Igor Zhukov _{3,

4} , Franciszek Kasprzykowski ₁ , Agnieszka Kubiś ₁ , Andrea D Lipińska ₂ , Sylwia Rodziewicz-Motowidło ₁

Affiliation

Herpesviruses are the most prevalent viruses that infect the human and animal body. They can escape a host immune response in numerous ways. One way is to block the TAP complex so that viral peptides, originating from proteasomal degradation, cannot be transported to the endoplasmic reticulum. As a result, a reduced number of MHC class I molecules appear on the surface of infected cells and, thus, the immune system is not efficiently activated. BoHV‐1‐encoded UL49.5 protein is one such TAP transporter inhibitor. This protein binds to TAP in such a way that its N‐terminal fragment interacts with the loops of the TAP complex, and the C‐terminus stimulates proteasomal degradation of TAP. Previous studies have indicated certain amino acid residues, especially the RRE(9–11) motif, within the helical structure of the UL49.5 N‐terminal fragment, as being crucial to the protein's activity. In this work, we investigated the effects of modifications within the RRE region on the spatial structure of the UL49.5 N‐terminal fragment. The introduced RRE(9–11) variations were designed to abolish or stabilize the structure of the α‐helix and, consequently, to increase or decrease protein activity compared to the wild type. The terminal structure of the peptides was established using circular dichroism (CD), 2D nuclear magnetic resonance (NMR), and molecular dynamics (MD) in membrane‐mimetic or membrane‐model environments. Our structural results show that in the RRE(9–11)AAA and E11G peptides the helical structure has been stabilized, whereas for the RRE(9–11)GGG peptide, as expected, the helix structure has partially unfolded compared to the native structure. These RRE modifications, in the context of the entire UL49.5 proteins, slightly altered their biological activity in human cells.

中文翻译：

RRE母题中的一级结构修饰对合成牛疱疹病毒1编码的UL49.5蛋白片段构象的影响的研究

疱疹病毒是感染人体和动物体的最普遍的病毒。它们可以多种方式逃脱宿主的免疫反应。一种方法是阻断TAP复合物，使源自蛋白酶体降解的病毒肽无法转运至内质网。结果，在感染的细胞表面上出现了数量减少的MHC I类分子，因此免疫系统没有得到有效激活。BoHV-1编码的UL49.5蛋白就是一种此类TAP转运蛋白抑制剂。该蛋白以其N末端片段与TAP复合物环相互作用的方式与TAP结合，而C末端刺激TAP的蛋白酶体降解。先前的研究表明，UL49.5 N端片段的螺旋结构中存在某些氨基酸残基，尤其是RRE（9-11）基序，对蛋白质的活性至关重要。在这项工作中，我们研究了RRE区域内修饰对UL49.5 N末端片段空间结构的影响。引入的RRE（9-11）变异旨在消除或稳定α-螺旋的结构，因此，与野生型相比，可以增加或减少蛋白质活性。在膜模拟或膜模型环境中，使用圆二色性（CD），二维核磁共振（NMR）和分子动力学（MD）确定了肽的末端结构。我们的结构结果表明，在RRE（9-11）AAA和E11G肽中，螺旋结构已稳定，而对于RRE（9-11）GGG肽，与预期结构相比，螺旋结构已部分展开。这些RRE修改，

更新日期：2021-02-12

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