Proteomic Characterization of 1000 Human and Murine Neutrophils Freshly Isolated From Blood and Sites of Sterile Inflammation.

IF 6.1 2区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Susmita Ghosh, Ali Ata Tuz, Martin Stenzel, Vikramjeet Singh, Mathis Richter, Oliver Soehnlein, Emanuel Lange, Robert Heyer, Zülal Cibir, Alexander Beer, Marcel Jung, Dennis Nagel, Dirk M Hermann, Anja Hasenberg, Anika Grüneboom, Albert Sickmann, Matthias Gunzer
{"title":"Proteomic Characterization of 1000 Human and Murine Neutrophils Freshly Isolated From Blood and Sites of Sterile Inflammation.","authors":"Susmita Ghosh, Ali Ata Tuz, Martin Stenzel, Vikramjeet Singh, Mathis Richter, Oliver Soehnlein, Emanuel Lange, Robert Heyer, Zülal Cibir, Alexander Beer, Marcel Jung, Dennis Nagel, Dirk M Hermann, Anja Hasenberg, Anika Grüneboom, Albert Sickmann, Matthias Gunzer","doi":"10.1016/j.mcpro.2024.100858","DOIUrl":null,"url":null,"abstract":"<p><p>Neutrophils are indispensable for defense against pathogens. Injured tissue-infiltrated neutrophils can establish a niche of chronic inflammation and promote degeneration. Studies investigated transcriptome of single-infiltrated neutrophils which could misinterpret molecular states of these post mitotic cells. However, neutrophil proteome characterization has been challenging due to low harvests from affected tissues. Here, we present a workflow to obtain proteome of 1000 murine and human tissue-infiltrated neutrophils. We generated spectral libraries containing ∼6200 mouse and ∼5300 human proteins from circulating neutrophils. 4800 mouse and 3400 human proteins were recovered from 1000 cells with 10<sup>2</sup>-10<sup>8</sup> copies/cell. Neutrophils from stroke-affected mouse brains adapted to the glucose-deprived environment with increased mitochondrial activity and ROS-production, while cells invading inflamed human oral cavities increased phagocytosis and granule release. We provide an extensive protein repository for resting human and mouse neutrophils, identify proteins lost in low input samples, thus enabling the proteomic characterization of limited tissue-infiltrated neutrophils.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100858"},"PeriodicalIF":6.1000,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular & Cellular Proteomics","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/j.mcpro.2024.100858","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0

Abstract

Neutrophils are indispensable for defense against pathogens. Injured tissue-infiltrated neutrophils can establish a niche of chronic inflammation and promote degeneration. Studies investigated transcriptome of single-infiltrated neutrophils which could misinterpret molecular states of these post mitotic cells. However, neutrophil proteome characterization has been challenging due to low harvests from affected tissues. Here, we present a workflow to obtain proteome of 1000 murine and human tissue-infiltrated neutrophils. We generated spectral libraries containing ∼6200 mouse and ∼5300 human proteins from circulating neutrophils. 4800 mouse and 3400 human proteins were recovered from 1000 cells with 102-108 copies/cell. Neutrophils from stroke-affected mouse brains adapted to the glucose-deprived environment with increased mitochondrial activity and ROS-production, while cells invading inflamed human oral cavities increased phagocytosis and granule release. We provide an extensive protein repository for resting human and mouse neutrophils, identify proteins lost in low input samples, thus enabling the proteomic characterization of limited tissue-infiltrated neutrophils.

从血液和无菌炎症部位新鲜分离的 1000 个人类和鼠类中性粒细胞的蛋白质组特征。
中性粒细胞是抵御病原体不可或缺的物质。损伤组织浸润的中性粒细胞可建立慢性炎症的生态位并促进退化。研究调查了单个浸润中性粒细胞的转录组,这可能会误解这些有丝分裂后细胞的分子状态。然而,由于从受影响组织中获取的蛋白质较少,嗜中性粒细胞蛋白质组的表征一直具有挑战性。在这里,我们介绍了一种获取 1000 个鼠和人组织浸润中性粒细胞蛋白质组的工作流程。我们从循环中性粒细胞中生成了包含 6,200 个小鼠蛋白质和 5,300 个人类蛋白质的谱库。从每细胞 102-108 个拷贝的 1000 个细胞中回收了 4,800 个小鼠蛋白质和 3,400 个人类蛋白质。受中风影响的小鼠大脑中的中性粒细胞适应了葡萄糖缺乏的环境,线粒体活性和 ROS 生成增加,而侵入发炎的人类口腔的细胞吞噬和颗粒释放增加。我们为静息的人和小鼠中性粒细胞提供了一个广泛的蛋白质库,识别了在低输入样本中丢失的蛋白质,从而实现了对有限组织浸润的中性粒细胞进行蛋白质组学表征。
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
Molecular & Cellular Proteomics
Molecular & Cellular Proteomics 生物-生化研究方法
CiteScore
11.50
自引率
4.30%
发文量
131
审稿时长
84 days
期刊介绍: The mission of MCP is to foster the development and applications of proteomics in both basic and translational research. MCP will publish manuscripts that report significant new biological or clinical discoveries underpinned by proteomic observations across all kingdoms of life. Manuscripts must define the biological roles played by the proteins investigated or their mechanisms of action. The journal also emphasizes articles that describe innovative new computational methods and technological advancements that will enable future discoveries. Manuscripts describing such approaches do not have to include a solution to a biological problem, but must demonstrate that the technology works as described, is reproducible and is appropriate to uncover yet unknown protein/proteome function or properties using relevant model systems or publicly available data. Scope: -Fundamental studies in biology, including integrative "omics" studies, that provide mechanistic insights -Novel experimental and computational technologies -Proteogenomic data integration and analysis that enable greater understanding of physiology and disease processes -Pathway and network analyses of signaling that focus on the roles of post-translational modifications -Studies of proteome dynamics and quality controls, and their roles in disease -Studies of evolutionary processes effecting proteome dynamics, quality and regulation -Chemical proteomics, including mechanisms of drug action -Proteomics of the immune system and antigen presentation/recognition -Microbiome proteomics, host-microbe and host-pathogen interactions, and their roles in health and disease -Clinical and translational studies of human diseases -Metabolomics to understand functional connections between genes, proteins and phenotypes
×
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