Impact of mutations in carbohydrate binding sites of tandem-repeat type galectin from Takifugu obscurus on its antimicrobial activity

IF 4.1 2区 农林科学 Q1 FISHERIES
Ying Huang , Li-Fan Cui , Rui Shen , Ding-Yi Chen , Min Jin , Xue Jiao , Yu-Guang Chen , Ming-Xuan Pan , Ya-Dong Hu , Zhe Zhao
{"title":"Impact of mutations in carbohydrate binding sites of tandem-repeat type galectin from Takifugu obscurus on its antimicrobial activity","authors":"Ying Huang ,&nbsp;Li-Fan Cui ,&nbsp;Rui Shen ,&nbsp;Ding-Yi Chen ,&nbsp;Min Jin ,&nbsp;Xue Jiao ,&nbsp;Yu-Guang Chen ,&nbsp;Ming-Xuan Pan ,&nbsp;Ya-Dong Hu ,&nbsp;Zhe Zhao","doi":"10.1016/j.fsi.2024.110018","DOIUrl":null,"url":null,"abstract":"<div><div>Galectins belong to a family of galactoside-binding proteins and exhibit diverse biological functions. In the present research, a tandem-repeat type galectin (named <em>ToGalectin</em>) was identified from obscure puffer <em>Takifugu obscurus</em>. The 296 amino acids ToGalectin contained two carbohydrate recognition domains (CRDs), one of which possessed two conserved carbohydrate binding motifs. Phylogenetic analysis showed that ToGalectin clustered tightly with other galectin-8 proteins from teleost fish. <em>ToGalectin</em> transcripts were ubiquitously expressed in all tissues examined and its expression was significantly upregulated in the liver, kidney, and intestine after <em>Vibrio harveyi</em> or <em>Staphylococcus aureus</em> infection. To investigate the effect of carbohydrate binding sites on biological activity, ToGalectin and its mutant (MUT-ToGalectin) were expressed and purified. The recombinant ToGalectin and MUT-ToGalectin proteins showed strong agglutinating activity against both <em>V</em>. <em>harveyi</em> and <em>S</em>. <em>aureus</em>. rToGalectin could bind to all tested carbohydrates and bacteria, whereas rMUT-ToGalectin bound to some carbohydrates and bacteria with specific and relatively strong affinity. rToGalectin significantly suppressed the growth of all six bacteria detected and promoted bacterial clearance <em>in vivo</em>, whereas MUT-ToGalectin inhibited the growth of only two bacterial species, which could be attributed to the differences in conserved motifs within the CRDs. Our results suggested that ToGalectin is involved in the immune response against bacterial infection and the clearance of pathogens in <em>T. obscurus</em>.</div></div>","PeriodicalId":12127,"journal":{"name":"Fish & shellfish immunology","volume":"155 ","pages":"Article 110018"},"PeriodicalIF":4.1000,"publicationDate":"2024-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Fish & shellfish immunology","FirstCategoryId":"97","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1050464824006636","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"FISHERIES","Score":null,"Total":0}
引用次数: 0

Abstract

Galectins belong to a family of galactoside-binding proteins and exhibit diverse biological functions. In the present research, a tandem-repeat type galectin (named ToGalectin) was identified from obscure puffer Takifugu obscurus. The 296 amino acids ToGalectin contained two carbohydrate recognition domains (CRDs), one of which possessed two conserved carbohydrate binding motifs. Phylogenetic analysis showed that ToGalectin clustered tightly with other galectin-8 proteins from teleost fish. ToGalectin transcripts were ubiquitously expressed in all tissues examined and its expression was significantly upregulated in the liver, kidney, and intestine after Vibrio harveyi or Staphylococcus aureus infection. To investigate the effect of carbohydrate binding sites on biological activity, ToGalectin and its mutant (MUT-ToGalectin) were expressed and purified. The recombinant ToGalectin and MUT-ToGalectin proteins showed strong agglutinating activity against both V. harveyi and S. aureus. rToGalectin could bind to all tested carbohydrates and bacteria, whereas rMUT-ToGalectin bound to some carbohydrates and bacteria with specific and relatively strong affinity. rToGalectin significantly suppressed the growth of all six bacteria detected and promoted bacterial clearance in vivo, whereas MUT-ToGalectin inhibited the growth of only two bacterial species, which could be attributed to the differences in conserved motifs within the CRDs. Our results suggested that ToGalectin is involved in the immune response against bacterial infection and the clearance of pathogens in T. obscurus.
钝角鲣鱼串联重复型半凝集素碳水化合物结合位点的突变对其抗菌活性的影响
半乳糖苷结合蛋白属于半乳糖苷结合蛋白家族,具有多种生物学功能。本研究从暗色河豚(Takifugu obscurus)中发现了一种串联重复型的半乳糖苷结合蛋白(ToGalectin)。296个氨基酸的ToGalectin含有两个碳水化合物识别域(CRD),其中一个具有两个保守的碳水化合物结合基序。系统进化分析表明,ToGalectin与其他来自远洋鱼类的galectin-8蛋白紧密聚类。ToGalectin转录本在所有受检组织中普遍表达,在哈维氏弧菌或金黄色葡萄球菌感染后,其在肝脏、肾脏和肠道中的表达显著上调。为了研究碳水化合物结合位点对生物活性的影响,我们表达并纯化了 ToGalectin 及其突变体(MUT-ToGalectin)。重组的 ToGalectin 和 MUT-ToGalectin 蛋白对 V. harveyi 和金黄色葡萄球菌都表现出很强的凝集活性。rToGalectin 能显著抑制所有六种检测到的细菌的生长,并促进细菌在体内的清除,而 MUT-ToGalectin 只能抑制两种细菌的生长,这可能是由于 CRDs 中保守基团的差异造成的。我们的研究结果表明,ToGalectin参与了钝口蟾对细菌感染的免疫反应和病原体的清除。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Fish & shellfish immunology
Fish & shellfish immunology 农林科学-海洋与淡水生物学
CiteScore
7.50
自引率
19.10%
发文量
750
审稿时长
68 days
期刊介绍: Fish and Shellfish Immunology rapidly publishes high-quality, peer-refereed contributions in the expanding fields of fish and shellfish immunology. It presents studies on the basic mechanisms of both the specific and non-specific defense systems, the cells, tissues, and humoral factors involved, their dependence on environmental and intrinsic factors, response to pathogens, response to vaccination, and applied studies on the development of specific vaccines for use in the aquaculture industry.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信