From sequence to function: Exploring biophysical properties of bacteriophage BFK20 lytic transglycosylase domain from the minor tail protein gp15

IF 2.5 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Kristina Papayova , Lucia Bocanova , Vladena Bauerova , Jacob Bauer , Nora Halgasova , Maria Kajsikova , Gabriela Bukovska
{"title":"From sequence to function: Exploring biophysical properties of bacteriophage BFK20 lytic transglycosylase domain from the minor tail protein gp15","authors":"Kristina Papayova ,&nbsp;Lucia Bocanova ,&nbsp;Vladena Bauerova ,&nbsp;Jacob Bauer ,&nbsp;Nora Halgasova ,&nbsp;Maria Kajsikova ,&nbsp;Gabriela Bukovska","doi":"10.1016/j.bbapap.2024.141044","DOIUrl":null,"url":null,"abstract":"<div><p>Bacteriophages have evolved different mechanisms of infection and penetration of bacterial cell walls. In <em>Siphoviridae</em>-like viruses, the inner tail proteins have a pivotal role in these processes and often encode lytic protein domains which increase infection efficiency. A soluble lytic transglycosylase (SLT) domain was identified in the minor tail protein gp15 from the BFK20 bacteriophage. Six fragments containing this SLT domain with adjacent regions of different lengths were cloned, expressed and purified. The biophysical properties of the two best expressing fragments were characterized by nanoDSF and CD spectroscopy, which showed that both fragments had a high refolding ability of 90 %. 3D modeling indicated that the bacteriophage BFK20 SLT domain is structurally similar to lysozyme. The degradation activity of these SLT proteins was evaluated using a lysozyme activity assay. BFK20 might use its transglycosylase activity to allow efficient phage DNA entry into the host cell by degrading bacterial peptidoglycan.</p></div>","PeriodicalId":8760,"journal":{"name":"Biochimica et biophysica acta. Proteins and proteomics","volume":null,"pages":null},"PeriodicalIF":2.5000,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biochimica et biophysica acta. Proteins and proteomics","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1570963924000517","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

Bacteriophages have evolved different mechanisms of infection and penetration of bacterial cell walls. In Siphoviridae-like viruses, the inner tail proteins have a pivotal role in these processes and often encode lytic protein domains which increase infection efficiency. A soluble lytic transglycosylase (SLT) domain was identified in the minor tail protein gp15 from the BFK20 bacteriophage. Six fragments containing this SLT domain with adjacent regions of different lengths were cloned, expressed and purified. The biophysical properties of the two best expressing fragments were characterized by nanoDSF and CD spectroscopy, which showed that both fragments had a high refolding ability of 90 %. 3D modeling indicated that the bacteriophage BFK20 SLT domain is structurally similar to lysozyme. The degradation activity of these SLT proteins was evaluated using a lysozyme activity assay. BFK20 might use its transglycosylase activity to allow efficient phage DNA entry into the host cell by degrading bacterial peptidoglycan.

Abstract Image

从序列到功能:探索噬菌体 BFK20 小尾蛋白 gp15 溶菌转糖基化酶结构域的生物物理特性。
噬菌体进化出了不同的感染和穿透细菌细胞壁的机制。在类Siphoviridae病毒中,内尾部蛋白在这些过程中起着关键作用,通常编码可提高感染效率的溶解蛋白结构域。在 BFK20 噬菌体的小尾蛋白 gp15 中发现了一个可溶性溶解性转糖基化酶(SLT)结构域。克隆、表达和纯化了含有该 SLT 结构域和不同长度相邻区域的六个片段。通过纳米脱硫荧光光谱和 CD 光谱对两个最佳表达片段的生物物理特性进行了表征,结果表明这两个片段的重折叠能力高达 90%。三维建模表明,噬菌体 BFK20 SLT 结构域与溶菌酶结构相似。利用溶菌酶活性测定法评估了这些 SLT 蛋白的降解活性。BFK20 可能利用其转糖基酶活性,通过降解细菌肽聚糖,使噬菌体 DNA 有效进入宿主细胞。
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
CiteScore
8.00
自引率
0.00%
发文量
55
审稿时长
33 days
期刊介绍: BBA Proteins and Proteomics covers protein structure conformation and dynamics; protein folding; protein-ligand interactions; enzyme mechanisms, models and kinetics; protein physical properties and spectroscopy; and proteomics and bioinformatics analyses of protein structure, protein function, or protein regulation.
×
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