增加分子量:使低温电镜结构测定低于100 kda的蛋白质

IF 2.7 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Koen Wentinck , Christos Gogou , Dimphna H. Meijer
{"title":"增加分子量:使低温电镜结构测定低于100 kda的蛋白质","authors":"Koen Wentinck ,&nbsp;Christos Gogou ,&nbsp;Dimphna H. Meijer","doi":"10.1016/j.crstbi.2022.09.005","DOIUrl":null,"url":null,"abstract":"<div><p>Significant advances in the past decade have enabled high-resolution structure determination of a vast variety of proteins by cryogenic electron microscopy single particle analysis. Despite improved sample preparation, next-generation imaging hardware, and advanced single particle analysis algorithms, small proteins remain elusive for reconstruction due to low signal-to-noise and lack of distinctive structural features. Multiple efforts have therefore been directed at the development of size-increase techniques for small proteins. Here we review the latest methods for increasing effective molecular weight of proteins &lt;100 ​kDa through target protein binding or target protein fusion - specifically by using nanobody-based assemblies, fusion tags, and symmetric scaffolds. Finally, we summarize these state-of-the-art techniques into a decision-tree to facilitate the design of tailored future approaches, and thus for further exploration of ever-smaller proteins that make up the largest part of the human genome.</p></div>","PeriodicalId":10870,"journal":{"name":"Current Research in Structural Biology","volume":"4 ","pages":"Pages 332-337"},"PeriodicalIF":2.7000,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9562432/pdf/","citationCount":"5","resultStr":"{\"title\":\"Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins\",\"authors\":\"Koen Wentinck ,&nbsp;Christos Gogou ,&nbsp;Dimphna H. Meijer\",\"doi\":\"10.1016/j.crstbi.2022.09.005\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Significant advances in the past decade have enabled high-resolution structure determination of a vast variety of proteins by cryogenic electron microscopy single particle analysis. Despite improved sample preparation, next-generation imaging hardware, and advanced single particle analysis algorithms, small proteins remain elusive for reconstruction due to low signal-to-noise and lack of distinctive structural features. Multiple efforts have therefore been directed at the development of size-increase techniques for small proteins. Here we review the latest methods for increasing effective molecular weight of proteins &lt;100 ​kDa through target protein binding or target protein fusion - specifically by using nanobody-based assemblies, fusion tags, and symmetric scaffolds. Finally, we summarize these state-of-the-art techniques into a decision-tree to facilitate the design of tailored future approaches, and thus for further exploration of ever-smaller proteins that make up the largest part of the human genome.</p></div>\",\"PeriodicalId\":10870,\"journal\":{\"name\":\"Current Research in Structural Biology\",\"volume\":\"4 \",\"pages\":\"Pages 332-337\"},\"PeriodicalIF\":2.7000,\"publicationDate\":\"2022-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9562432/pdf/\",\"citationCount\":\"5\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current Research in Structural Biology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2665928X22000290\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Research in Structural Biology","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2665928X22000290","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 5

摘要

在过去的十年中,通过低温电子显微镜单粒子分析,可以对多种蛋白质进行高分辨率结构测定。尽管改进了样品制备,下一代成像硬件和先进的单颗粒分析算法,但由于低信噪比和缺乏独特的结构特征,小蛋白质仍然难以重建。因此,多种努力都是针对小蛋白质的尺寸增加技术的发展。在这里,我们回顾了通过靶蛋白结合或靶蛋白融合(特别是使用基于纳米体的组件、融合标签和对称支架)来增加蛋白质有效分子量(100 kDa)的最新方法。最后,我们将这些最先进的技术总结为一个决策树,以促进设计量身定制的未来方法,从而进一步探索构成人类基因组最大部分的更小的蛋白质。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins

Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins

Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins

Putting on molecular weight: Enabling cryo-EM structure determination of sub-100-kDa proteins

Significant advances in the past decade have enabled high-resolution structure determination of a vast variety of proteins by cryogenic electron microscopy single particle analysis. Despite improved sample preparation, next-generation imaging hardware, and advanced single particle analysis algorithms, small proteins remain elusive for reconstruction due to low signal-to-noise and lack of distinctive structural features. Multiple efforts have therefore been directed at the development of size-increase techniques for small proteins. Here we review the latest methods for increasing effective molecular weight of proteins <100 ​kDa through target protein binding or target protein fusion - specifically by using nanobody-based assemblies, fusion tags, and symmetric scaffolds. Finally, we summarize these state-of-the-art techniques into a decision-tree to facilitate the design of tailored future approaches, and thus for further exploration of ever-smaller proteins that make up the largest part of the human genome.

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
CiteScore
4.60
自引率
0.00%
发文量
33
审稿时长
104 days
×
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