食品与发酵工业 >

2023 , Vol. 49 >Issue 13: 144 - 151

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.035099

研究报告

分子对接技术筛选菲律宾蛤仔抗炎肽

  • 姜玉 ,
  • 林海生 ,
  • 秦小明 ,
  • 高加龙 ,
  • 曹文红 ,
  • 陈忠琴 ,
  • 郑惠娜 ,
  • 伍彬
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  • 1(广东海洋大学 食品科技学院,广东 湛江,524088)
    2(广东省水产品加工与安全重点实验室,广东普通高等学校水产品深加工重点实验室,国家贝类加工技术研发分中心 (湛江),南海生物资源开发与利用协同创新中心,广东 湛江,524088)
第一作者:硕士研究生(林海生博士为通信作者,E-mail:haishenglin@163.com)

收稿日期: 2023-02-11

  修回日期: 2023-03-14

  网络出版日期: 2023-08-07

基金资助

财政部和农业农村部国家现代农业产业技术体系资助项目(CARS-49);广东海洋大学博士启动项目(R17082);广东省现代农业产业技术体系创新团队项目(2021KJ146);广东省高等学校科技创新团队项目(2021KCXTD021)

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Screening of anti-inflammatory peptides from Ruditapes philippinarum by molecular docking

  • JIANG Yu ,
  • LIN Haisheng ,
  • QIN Xiaoming ,
  • GAO Jialong ,
  • CAO Wenhong ,
  • CHEN Zhongqin ,
  • ZHENG Huina ,
  • WU Bin
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  • 1(College of Food Science and Technology, Guangdong Ocean University, Zhanjiang 524088, China)
    2(Guangdong Key Laboratory of Aquatic Products Processing and Safety, Key Laboratory of Aquatic Products Deep Processing of Guangdong Universities, National Research and Development Branch Center for Shellfish Processing Technology (Zhanjiang), South China Sea Biological Resources Development and Utilization Collaborative Innovation Center, Zhanjiang 524088, China)

Received date: 2023-02-11

  Revised date: 2023-03-14

  Online published: 2023-08-07

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摘要

基于分子对接技术筛选菲律宾蛤仔(Ruditapes philippinarum)来源的具有抗炎作用的活性肽。采用三氯乙酸-丙酮沉淀法提取菲律宾蛤仔蛋白质,通过双向凝胶电泳(2-dimensional electro phoresis,2-DE)技术和液质联用(LC-MS/MS)技术分离和鉴定其蛋白质,借助BIOPEP-UWM在线工具对主要蛋白进行虚拟酶解,使用PeptideRanker和ToxinPred等程序对肽段进行评分和理化性质分析;利用Discovery studio 2019和AutoDock Vina软件进行分子对接筛选潜在抗炎肽片段,探讨人工合成抗炎肽对LPS诱导的RAW264.7细胞炎症的改善作用。结果表明,原肌球蛋白和组蛋白为肌肉组织的主要蛋白质。选用原肌球蛋白作为底物,通过虚拟酶解和分子对接筛选出8种潜在的新颖活性肽。其中DQTF与TLR2和TLR4分别存在 7个氨基酸残基和8个氨基酸残基结合位点,而GYTR与TLR2分子中的10个氨基酸残基和TLR4分子中的7个氨基酸残基紧密结合(结合能均低于-5 kcal/mol),表明其是潜在的抗炎肽。活性验证结果表明,2条合成寡肽均能显著增强RAW264.7细胞活力(P＜0.001),并显著降低细胞产NO能力(P＜0.05),表现出良好的抗炎活性。综上,基于虚拟酶解和分子对接方法可用于筛选菲律宾蛤仔来源的抗炎肽。

关键词： 菲律宾蛤仔; 原肌球蛋白; 虚拟酶解; 分子对接; RAW264.7细胞; 抗炎寡肽

本文引用格式

姜玉 , 林海生 , 秦小明 , 高加龙 , 曹文红 , 陈忠琴 , 郑惠娜 , 伍彬 . 分子对接技术筛选菲律宾蛤仔抗炎肽[J]. 食品与发酵工业, 2023 , 49(13) : 144 -151 . DOI: 10.13995/j.cnki.11-1802/ts.035099

Abstract

Screening of active peptides with anti-inflammatory effects of Ruditapes philippinarum based on molecular docking. In this study, the proteins of R. philippinarum were extracted by trichloroacetic acid-acetone precipitation and identified by 2-DE and LC-MS/MS. The main protein was enzymolized by BIOPEP-UWM and the PeptideRanker and ToxinPred programs were used to evaluate and predict the physicochemical properties of peptides. Molecular docking was performed using Discovery studio 2019 and AutoDock Vina to screen potential anti-inflammatory peptide fragments, the improvement of LPS-induced inflammation in RAW264.7 cells was verified by synthetic anti-inflammatory peptides. The results showed that tropomyosin and histone were the main proteins of muscle tissue. Eight potential unreported active peptides were screened to use tropomyosin as substrate by virtual enzymolysis. DQTF had seven amino acid residues and eight amino acid residues binding sites with TLR2 and TLR4 respectively, while GYTR was closely combined with ten amino acid residues in TLR2 molecule and seven amino acid residues in TLR4 molecule , with the binding energy lower than -5 kcal/mol, indicating that it was a potential anti-inflammatory peptide. The activity verification results showed that the two synthetic oligopeptides could significantly enhance the cell viability (P<0.001) and reduce the ability of NO production(P<0.05), showing great anti-inflammatory activity. In conclusion, virtual enzymolysis and molecular docking methods can be used to screen anti-inflammatory peptides derived from R. philippinarum.

Key words： Ruditapes philippinarum; tropomyosin; virtual enzymatic hydrolysis; molecular docking; RAW 264.7 cells; anti-inflammatory oligopeptides

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