利用植物胱抑素的功能多样性设计对草食性节肢动物消化蛋白酶具有高活性的抑制剂变体。

IF 5.5 2区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
FEBS Journal Pub Date : 2022-04-01 Epub Date: 2021-12-01 DOI:10.1111/febs.16288
Jonathan Tremblay, Marie-Claire Goulet, Juan Vorster, Charles Goulet, Dominique Michaud
{"title":"利用植物胱抑素的功能多样性设计对草食性节肢动物消化蛋白酶具有高活性的抑制剂变体。","authors":"Jonathan Tremblay,&nbsp;Marie-Claire Goulet,&nbsp;Juan Vorster,&nbsp;Charles Goulet,&nbsp;Dominique Michaud","doi":"10.1111/febs.16288","DOIUrl":null,"url":null,"abstract":"<p><p>Protein engineering approaches have been proposed to improve the inhibitory properties of plant cystatins against herbivorous arthropod digestive proteases, generally involving the site-directed mutagenesis of functionally relevant amino acids or the selection of improved inhibitor variants by phage display approaches. Here, we propose a novel approach where the function-related structural elements of a cystatin are substituted by the corresponding elements of an alternative cystatin. Inhibitory assays were first performed with 20 representative plant cystatins and model Cys proteases, including arthropod proteases, to appreciate the extent of functional variability among the plant cystatin family. The most, and less, potent of these cystatins were then used as 'donors' of structural elements to create hybrids of tomato cystatin SlCYS8 used as a model 'recipient' inhibitor. In brief, inhibitory activities against Cys proteases strongly differed from one plant cystatin to another, with K<sub>i (papain)</sub> values diverging by more than 30-fold and inhibitory rates against arthropod proteases varying by up to 50-fold depending on the enzymes assessed. In line with theoretical assumptions from docking models generated for different Cys protease-cystatin combinations, structural element substitutions had a strong impact on the activity of recipient cystatin SlCYS8, positive or negative depending on the basic inhibitory potency of the donor cystatin. Our data confirm the wide variety of cystatin inhibitory profiles among plant taxa. They also demonstrate the usefulness of these proteins as a pool of discrete structural elements for the design of cystatin variants with improved potency against herbivorous pest digestive Cys proteases.</p>","PeriodicalId":12261,"journal":{"name":"FEBS Journal","volume":"289 7","pages":"1827-1841"},"PeriodicalIF":5.5000,"publicationDate":"2022-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"6","resultStr":"{\"title\":\"Harnessing the functional diversity of plant cystatins to design inhibitor variants highly active against herbivorous arthropod digestive proteases.\",\"authors\":\"Jonathan Tremblay,&nbsp;Marie-Claire Goulet,&nbsp;Juan Vorster,&nbsp;Charles Goulet,&nbsp;Dominique Michaud\",\"doi\":\"10.1111/febs.16288\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Protein engineering approaches have been proposed to improve the inhibitory properties of plant cystatins against herbivorous arthropod digestive proteases, generally involving the site-directed mutagenesis of functionally relevant amino acids or the selection of improved inhibitor variants by phage display approaches. Here, we propose a novel approach where the function-related structural elements of a cystatin are substituted by the corresponding elements of an alternative cystatin. Inhibitory assays were first performed with 20 representative plant cystatins and model Cys proteases, including arthropod proteases, to appreciate the extent of functional variability among the plant cystatin family. The most, and less, potent of these cystatins were then used as 'donors' of structural elements to create hybrids of tomato cystatin SlCYS8 used as a model 'recipient' inhibitor. In brief, inhibitory activities against Cys proteases strongly differed from one plant cystatin to another, with K<sub>i (papain)</sub> values diverging by more than 30-fold and inhibitory rates against arthropod proteases varying by up to 50-fold depending on the enzymes assessed. In line with theoretical assumptions from docking models generated for different Cys protease-cystatin combinations, structural element substitutions had a strong impact on the activity of recipient cystatin SlCYS8, positive or negative depending on the basic inhibitory potency of the donor cystatin. Our data confirm the wide variety of cystatin inhibitory profiles among plant taxa. They also demonstrate the usefulness of these proteins as a pool of discrete structural elements for the design of cystatin variants with improved potency against herbivorous pest digestive Cys proteases.</p>\",\"PeriodicalId\":12261,\"journal\":{\"name\":\"FEBS Journal\",\"volume\":\"289 7\",\"pages\":\"1827-1841\"},\"PeriodicalIF\":5.5000,\"publicationDate\":\"2022-04-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"6\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"FEBS Journal\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1111/febs.16288\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2021/12/1 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q1\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"FEBS Journal","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1111/febs.16288","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2021/12/1 0:00:00","PubModel":"Epub","JCR":"Q1","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 6

摘要

已经提出了蛋白质工程方法来提高植物胱抑素对食草节肢动物消化蛋白酶的抑制性能,通常涉及功能相关氨基酸的定点诱变或通过噬菌体展示方法选择改进的抑制剂变体。在这里,我们提出了一种新的方法,其中胱抑素的功能相关结构元件被替代胱抑素的相应元件所取代。首先对20种具有代表性的植物胱抑素和模型胱抑素蛋白酶(包括节肢动物蛋白酶)进行抑制试验,以了解植物胱抑素家族之间功能差异的程度。然后,这些半胱抑素中最有效和最不有效的半胱抑素被用作结构元件的“供体”,以创造西红柿半胱抑素SlCYS8的杂交体,作为模型“受体”抑制剂。简而言之,不同植物的胱抑素对Cys蛋白酶的抑制活性差异很大,Ki(木瓜蛋白酶)值相差超过30倍,节肢动物蛋白酶的抑制率相差高达50倍,这取决于所评估的酶。根据对不同Cys蛋白酶-胱抑素组合的对接模型的理论假设,结构元件取代对受体胱抑素SlCYS8的活性有很强的影响,根据供体胱抑素的基本抑制效力,影响是正的还是负的。我们的数据证实了植物类群中胱抑素抑制谱的广泛多样性。他们还证明了这些蛋白质作为一个离散结构元素池的有用性,用于设计具有提高抗草食性害虫消化Cys蛋白酶效力的胱抑素变体。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Harnessing the functional diversity of plant cystatins to design inhibitor variants highly active against herbivorous arthropod digestive proteases.

Protein engineering approaches have been proposed to improve the inhibitory properties of plant cystatins against herbivorous arthropod digestive proteases, generally involving the site-directed mutagenesis of functionally relevant amino acids or the selection of improved inhibitor variants by phage display approaches. Here, we propose a novel approach where the function-related structural elements of a cystatin are substituted by the corresponding elements of an alternative cystatin. Inhibitory assays were first performed with 20 representative plant cystatins and model Cys proteases, including arthropod proteases, to appreciate the extent of functional variability among the plant cystatin family. The most, and less, potent of these cystatins were then used as 'donors' of structural elements to create hybrids of tomato cystatin SlCYS8 used as a model 'recipient' inhibitor. In brief, inhibitory activities against Cys proteases strongly differed from one plant cystatin to another, with Ki (papain) values diverging by more than 30-fold and inhibitory rates against arthropod proteases varying by up to 50-fold depending on the enzymes assessed. In line with theoretical assumptions from docking models generated for different Cys protease-cystatin combinations, structural element substitutions had a strong impact on the activity of recipient cystatin SlCYS8, positive or negative depending on the basic inhibitory potency of the donor cystatin. Our data confirm the wide variety of cystatin inhibitory profiles among plant taxa. They also demonstrate the usefulness of these proteins as a pool of discrete structural elements for the design of cystatin variants with improved potency against herbivorous pest digestive Cys proteases.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
FEBS Journal
FEBS Journal 生物-生化与分子生物学
CiteScore
11.70
自引率
1.90%
发文量
375
审稿时长
1 months
期刊介绍: The FEBS Journal is an international journal devoted to the rapid publication of full-length papers covering a wide range of topics in any area of the molecular life sciences. The criteria for acceptance are originality and high quality research, which will provide novel perspectives in a specific area of research, and will be of interest to our broad readership. The journal does not accept papers that describe the expression of specific genes and proteins or test the effect of a drug or reagent, without presenting any biological significance. Papers describing bioinformatics, modelling or structural studies of specific systems or molecules should include experimental data.
×
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