与葡萄糖结合的双色 Dioclea Lectin (DBL):纯化、表征、结构分析和抗菌特性

IF 1.9 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY The Protein Journal Pub Date : 2024-04-14 DOI:10.1007/s10930-024-10199-9
Willian F. Reis, Marcos E. S. Silva, Ana C. S. Gondim, Renato C. F. Torres, Rômulo F. Carneiro, Celso S. Nagano, Alexandre H. Sampaio, Claudener S. Teixeira, Lenita C. B. F. Gomes, Bruno L. Sousa, Alexandre L. Andrade, Edson H. Teixeira, Mayron A. Vasconcelos
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引用次数: 0

摘要

在这项研究中,我们通过亲和纯化法纯化了从Dioclea bicolor种子中分离出来的一种凝集素(DBL)。电泳分析显示,DBL 有三条带,分别为 α、β 和 γ 链,分子质量分别约为 29、14 和 12 kDa。凝胶过滤色谱法显示,原生形式的 DBL 分子质量约为 100 kDa,表明它是一个四聚体。有趣的是,DBL 诱导的血凝作用受到几种苷类、甘露苷类、氨苄西林和四环素的抑制,最低抑制浓度(MIC)值为 1.56-50 mM。对 DBL 完整氨基酸序列的分析表明,DBL 含有 237 个氨基酸,与其他 Diocleinae 凝集素高度相似。圆二色性显示了DBL突出的β-片状二级结构。此外,DBL结构预测显示离散优化蛋白能量(DOPE)得分为-26,642.69141/归一化DOPE得分为-1.84041。根据其三维结构发现,DBL单体由β-三明治组成。分子对接显示了DBL与α-D-葡萄糖、N-乙酰-D-葡萄糖胺、α-D-甘露糖、α-甲基-D-甘露糖苷、氨苄西林和四环素之间的相互作用。此外,DBL 还具有抗菌活性,其 MIC 值为 125 μg/mL,与氨苄西林和四环素联用可产生协同效应(部分抑制浓度指数≤0.5)。此外,DBL 还能明显抑制生物膜的形成,且对小鼠成纤维细胞无毒性(p < 0.05)。这些结果表明,DBL 具有抗菌活性,并能与抗生素协同作用。
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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Glucose-Binding Dioclea bicolor Lectin (DBL): Purification, Characterization, Structural Analysis, and Antibacterial Properties

In this study, we purified a lectin isolated from the seeds of Dioclea bicolor (DBL) via affinity purification. Electrophoresis analysis revealed that DBL had three bands, α, β, and γ chains, with molecular masses of approximately 29, 14, and 12 kDa, respectively. Gel filtration chromatography revealed that the native form of DBL had a molecular mass of approximately 100 kDa, indicating that it is a tetramer. Interestingly, DBL-induced hemagglutination was inhibited by several glucosides, mannosides, ampicillin, and tetracycline with minimum inhibitory concentration (MIC) values of 1.56–50 mM. Analysis of the complete amino acid sequence of DBL revealed the presence of 237 amino acids with high similarity to other Diocleinae lectins. Circular dichroism showed the prominent β-sheet secondary structure of DBL. Furthermore, DBL structure prediction revealed a Discrete Optimized Protein Energy (DOPE) score of –26,642.69141/Normalized DOPE score of –1.84041. The DBL monomer was found to consist a β-sandwich based on its 3D structure. Molecular docking showed the interactions between DBL and α-D-glucose, N-acetyl-D-glucosamine, α-D-mannose, α-methyl-D-mannoside, ampicillin, and tetracycline. In addition, DBL showed antimicrobial activity with an MIC of 125 μg/mL and exerted synergistic effects in combination with ampicillin and tetracycline (fractional inhibitory concentration index ≤ 0.5). Additionally, DBL significantly inhibited biofilm formation and showed no toxicity in murine fibroblasts (p < 0.05). These results suggest that DBL exhibits antimicrobial activity and works synergistically with antibiotics.

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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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