• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Molecular Biology >Specific Recognition of a Single-Stranded RNA Sequence by a Synthetic Antibody Fragment
【24h】

Specific Recognition of a Single-Stranded RNA Sequence by a Synthetic Antibody Fragment

机译：合成抗体片段对单链RNA序列的特异性识别。

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

Antibodies that bind RNA represent an unrealized source of reagents for synthetic biology and for characterizing cellular transcriptomes. However, facile access to RNA-binding antibodies requires the engineering of effective Fab libraries guided by the knowledge of the principles that govern RNA recognition. Here, we describe a Fab identified from a minimalist synthetic library during phage display against a branched RNA target. The Fab (BRG) binds with 20 nM dissociation constant to a single-stranded RNA (ssRNA) sequence adjacent to the branch site and can block the action of debranchase enzyme. We report the crystal structure in complex with RNA target at 2.38 angstrom. The Fab traps the RNA in a hairpin conformation that contains a 2-bp duplex capped by a tetraloop. The paratope surface consists of residues located in four complementarity-determining regions including a major contribution from H3, which adopts a helical structure that projects into a deep, wide groove formed by the RNA. The amino acid composition of the paratope reflects the library diversity, consisting mostly of tyrosine and serine residues and a small but significant contribution from a single arginine residue. This structure, involving the recognition of ssRNA via a stem-loop conformation, together with our two previous structures involving the recognition of an RNA hairpin loop and an RNA tertiary structure, reveals the capacity of minimalist libraries biased with tyrosine, serine, glycine, and arginine to form binding surfaces for specific RNA conformations and distinct levels of RNA structural hierarchy. (C) 2016 Elsevier Ltd. All rights reserved.
机译：结合RNA的抗体代表了用于合成生物学和表征细胞转录组的试剂的未实现来源。然而，要想容易地获得RNA结合抗体，就需要对有效的Fab文库进行改造，并以控制RNA识别的原理为指导。在这里，我们描述了在针对分支RNA靶标的噬菌体展示过程中从简约的合成文库中识别出的Fab。 Fab（BRG）以20 nM的解离常数与邻近分支位点的单链RNA（ssRNA）序列结合，并可以阻断脱支酶的作用。我们报告与2.38埃的RNA目标复杂的晶体结构。 Fab以发夹构型捕获RNA，该构型包含一个被四环封端的2 bp双链体。互补位表面由位于四个互补决定区的残基组成，包括H3的主要贡献，H3采用螺旋结构，该结构伸入RNA形成的深而宽的凹槽中。互补位的氨基酸组成反映了文库的多样性，主要由酪氨酸和丝氨酸残基组成，而单个精氨酸残基的贡献很小但很重要。这种结构涉及通过茎环构象识别ssRNA，再加上我们先前涉及的两个涉及RNA发夹环和RNA三级结构识别的结构，揭示了被酪氨酸，丝氨酸，甘氨酸和精氨酸形成用于特定RNA构象和不同水平的RNA结构层次的结合表面。 （C）2016 Elsevier Ltd.保留所有权利。

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