变形链球菌粘附素P1的C末端结构域的骨干NMR共振归属。

IF 0.8 4区 生物学 Q4 BIOPHYSICS
Emily-Qingqing Peng, M. Luiza Caldas Nogueira, Gwladys Rivière, L. Jeannine Brady, Joanna R. Long
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引用次数: 0

摘要

粘附素P1(又名AgI/II)在介导变异链球菌在口腔中的附着以及调节生物膜的发育和成熟中起着关键作用。P1的天然截短产物抗原II(AgII)在细菌生命周期内采用可溶性、单体和不溶性淀粉样蛋白生成形式。单体参与重要的四元相互作用,促进细胞粘附,功能性淀粉样蛋白形式促进成熟生物膜的分离。异源的51kD C123构建体包含大部分AgII,并且先前通过X射线晶体学进行了表征。C123包含三个结构同源结构域,C1、C2和C3。使用原始C123构建体或其C3结构域制备的NMR样品产生中等分辨率的NMR光谱。使用Alphabold,我们重新分析了P1序列,以更好地识别C123的结构域边界,特别是C3结构域。然后,我们生成了一个更易于处理的结构体,用于单体形式的NMR研究,包括与其他蛋白质的四元相互作用。相对于先前的构建体,在C末端添加7个氨基酸大大改善了C3的光谱色散。在这里,我们报道了新结构的骨架核磁共振分配,并表征了它的一些四元相互作用。这些数据与Alphabold预测的结构非常一致,对于缺乏七个C末端氨基酸的构建体,与C123晶体结构中的C3结构域相比,Alphabold包含额外的β-片二级结构。其与已知蛋白质伴侣的四元相互作用与先前的竞争性结合测定非常一致。该构建体可用于进一步的NMR研究,包括蛋白质-蛋白质相互作用研究,并评估环境条件对C123从单体转变为淀粉样蛋白形式时C3结构和动力学的影响。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Backbone NMR resonance assignments for the C terminal domain of the Streptococcus mutans adhesin P1

Backbone NMR resonance assignments for the C terminal domain of the Streptococcus mutans adhesin P1

Adhesin P1 (aka AgI/II) plays a pivotal role in mediating Streptococcus mutans attachment in the oral cavity, as well as in regulating biofilm development and maturation. P1’s naturally occurring truncation product, Antigen II (AgII), adopts both soluble, monomeric and insoluble, amyloidogenic forms within the bacterial life cycle. Monomers are involved in important quaternary interactions that promote cell adhesion and the functional amyloid form promotes detachment of mature biofilms. The heterologous, 51-kD C123 construct comprises most of AgII and was previously characterized by X-ray crystallography. C123 contains three structurally homologous domains, C1, C2, and C3. NMR samples made using the original C123 construct, or its C3 domain, yielded moderately resolved NMR spectra. Using Alphafold, we re-analyzed the P1 sequence to better identify domain boundaries for C123, and in particular the C3 domain. We then generated a more tractable construct for NMR studies of the monomeric form, including quaternary interactions with other proteins. The addition of seven amino acids at the C-terminus greatly improved the spectral dispersion for C3 relative to the prior construct. Here we report the backbone NMR resonance assignments for the new construct and characterize some of its quaternary interactions. These data are in good agreement with the structure predicted by Alphafold, which contains additional β-sheet secondary structure compared to the C3 domain in the C123 crystal structure for a construct lacking the seven C-terminal amino acids. Its quaternary interactions with known protein partners are in good agreement with prior competitive binding assays. This construct can be used for further NMR studies, including protein-protein interaction studies and assessing the impact of environmental conditions on C3 structure and dynamics within C123 as it transitions from monomer to amyloid form.

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来源期刊
Biomolecular NMR Assignments
Biomolecular NMR Assignments 生物-光谱学
CiteScore
1.70
自引率
11.10%
发文量
59
审稿时长
6-12 weeks
期刊介绍: Biomolecular NMR Assignments provides a forum for publishing sequence-specific resonance assignments for proteins and nucleic acids as Assignment Notes. Chemical shifts for NMR-active nuclei in macromolecules contain detailed information on molecular conformation and properties. Publication of resonance assignments in Biomolecular NMR Assignments ensures that these data are deposited into a public database at BioMagResBank (BMRB; http://www.bmrb.wisc.edu/), where they are available to other researchers. Coverage includes proteins and nucleic acids; Assignment Notes are processed for rapid online publication and are published in biannual online editions in June and December.
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