与载体共轭有助于揭示 Parvovirus B19 磷脂酶结构域肽的酸化诱导结构转变。

IF 1.9 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY The Protein Journal Pub Date : 2024-07-09 DOI:10.1007/s10930-024-10209-w
Vladislav Victorovich Khrustalev, Olga Victorovna Khrustaleva, Aleksander Nicolaevich Stojarov, Anastasia Aleksandrovna Akunevich, Oleg Evgenyevich Baranov, Anna Vladimirovna Popinako, Elena Olegovna Samoilovich, Marina Anatolyevna Yermolovich, Galina Valeryevna Semeiko, Victoria Igorevna Cheprasova, Egor Gennadyevich Sapon, Nikolai Vladimirovich Shalygo, Victor Vitoldovich Poboinev, Tatyana Aleksandrovna Khrustaleva, Bahdan Vyacheslavovich Ranishenka, Ulyana Vitalyevna Kharytonova, Daniel Bush
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引用次数: 0

摘要

由于短肽在水生缓冲液中容易形成寡聚体,因此对大型蛋白质的结构域和肽进行光谱研究非常复杂,但将肽与载体蛋白共轭可能会有所帮助。在这项研究中,我们证实了 VP1 Parvovirus B19 包膜蛋白磷脂酶 A2 结构域的 SK30 肽片段(残基:144-159;164;171-183;序列:SAVDSAARIHDFR)可以与载体蛋白结合:根据 CD 光谱结果,只有在与 BSA(通过额外的 N 端 Cys 残基将其转化为 CSK31 肽和 SMCC 连接器)共轭的情况下,该肽才会在酸性介质中从随机线圈变为α螺旋。与此相反,根据红外光谱、CD 光谱、蓝色原生凝胶电泳和离心超滤的结果,未共轭的 SK30 肽不会发生这种转变,因为它形成了稳定的寡聚体,由分子间反平行的 beta 片层连接起来,VP1 蛋白的整个分离磷脂酶结构域可能也是如此。然而,磷脂酶结构域作为长 VP1 荚膜蛋白的一部分,在内溶酶体介质酸化过程中可能会通过质子化的 His153 和 Asp175 之间形成接触的方式改变其折叠,从而促进其 N 端部分从无规线圈转变为α螺旋。这项研究为疫苗开发开辟了前景,因为针对 CSK31 多肽与 BSA 共轭物的兔多克隆抗体(其中磷脂酶 A2 结构域的第二个α螺旋的结构应再现)可与 VP1 Parvovirus B19 病毒荚膜的完整重组独特部分(残基:1-227)的表位结合。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Conjugation with the Carrier Helped to Reveal acidification-Induced Structural Shift in the Peptide from Phospholipase Domain of Parvovirus B19

Spectroscopic studies on domains and peptides of large proteins are complicated because of the tendency of short peptides to form oligomers in aquatic buffers, but conjugation of a peptide with a carrier protein may be helpful. In this study we approved that a fragment of SK30 peptide from phospholipase A2 domain of VP1 Parvovirus B19 capsid protein (residues: 144–159; 164; 171–183; sequence: SAVDSAARIHDFRYSQLAKLGINPYTHWTVADEELLKNIK) turns from random coil to alpha helix in the acidic medium only in case if it had been conjugated with BSA (through additional N-terminal Cys residue, turning it into CSK31 peptide, and SMCC linker) according to CD-spectroscopy results. In contrast, unconjugated SK30 peptide does not undergo such shift because it forms stable oligomers connected by intermolecular antiparallel beta sheet, according to IR-spectroscopy, CD-spectroscopy, blue native gel electrophoresis and centrifugal ultrafiltration, as, probably, the whole isolated phospholipase domain of VP1 protein does. However, being a part of the long VP1 capsid protein, phospholipase domain may change its fold during the acidification of the medium in the endolysosome by the way of the formation of contacts between protonated His153 and Asp175, promoting the shift from random coil to alpha helix in its N-terminal part. This study opens up a perspective of vaccine development, since rabbit polyclonal antibodies against the conjugate of CSK31 peptide with BSA, in which the structure of the second alpha helix from the phospholipase A2 domain should be reproduced, can bind epitopes of the complete recombinant unique part of VP1 Parvovirus B19 capsid (residues: 1-227).

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来源期刊
The Protein Journal
The Protein Journal 生物-生化与分子生物学
CiteScore
5.20
自引率
0.00%
发文量
57
审稿时长
12 months
期刊介绍: The Protein Journal (formerly the Journal of Protein Chemistry) publishes original research work on all aspects of proteins and peptides. These include studies concerned with covalent or three-dimensional structure determination (X-ray, NMR, cryoEM, EPR/ESR, optical methods, etc.), computational aspects of protein structure and function, protein folding and misfolding, assembly, genetics, evolution, proteomics, molecular biology, protein engineering, protein nanotechnology, protein purification and analysis and peptide synthesis, as well as the elucidation and interpretation of the molecular bases of biological activities of proteins and peptides. We accept original research papers, reviews, mini-reviews, hypotheses, opinion papers, and letters to the editor.
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