Time-of-Flight Fragmentation Spectra Generated by the Proteomic Analysis of Single Human Cells Do Not Exhibit Atypical Fragmentation Patterns.
Time-of-Flight Fragmentation Spectra Generated by the Proteomic Analysis of Single Human Cells Do Not Exhibit Atypical Fragmentation Patterns.
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DOI：
10.1021/acs.jproteome.2c00715
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发表时间：
2023-03-03
影响因子：
4.4
通讯作者：
Orsburn, Benjamin C.
中科院分区：
文献类型：
--
作者：
Orsburn, Benjamin C.
最近的工作详细描述了来自单个人类细胞的多肽产生的碎裂光谱的独特特征。这份有价值的报告利用超高场轨道陷阱，并直接比较了从高浓度体细胞HeLa裂解物获得的光谱与从相同样品的纳克稀释物获得的光谱以及从纳米孔处理的单个HeLa细胞获得的光谱。分析表明，在高电池负载和单电池负载下产生的碎裂光谱之间存在显著差异，最显著的是，高质量Y系列碎片离子的损失。由于Orbitrap和飞行时间质量分析仪在物理上存在显著差异，似乎有必要进行比较。用分离的单个胰腺癌细胞与由100个细胞组成的细胞池进行了类似的分析。虽然对之前的Orbitrap数据的重新分析支持了作者的原始发现，但在来自单个人类细胞的多肽的飞行时间质谱图中没有观察到同样的趋势。当直接比较在相同质量分析仪上产生的批量数据和单个细胞数据之间的匹配强度碎片值时，结果尤其显著。仪器采集文件、处理后的数据和谱库可通过ACCESS MSV000090635在MASS上公开获得。
Recent work detailed the unique characteristics of fragmentation spectra derived from peptides from single human cells. This valuable report utilized an ultrahigh-field Orbitrap and directly compared the spectra obtained from high-concentration bulk cell HeLa lysates to those obtained from nanogram dilutions of the same and from nanowell-processed single HeLa cells. The analysis demonstrated marked differences between the fragmentation spectra generated at high and single-cell loads, most strikingly, the loss of high-mass y-series fragment ions. As significant differences exist in the physics of Orbitrap and time-of-flight mass analyzers, a comparison appeared warranted. A similar analysis was performed using isolated single pancreatic cancer cells compared to pools consisting of 100 cells. While a reanalysis of the prior Orbitrap data supports the author’s original findings, the same trends are not observed in time-of-flight mass spectra of peptides from single human cells. The results are particularly striking when directly comparing the matched intensity fragment values between bulk and single-cell data generated on the same mass analyzers. Instrument acquisition files, processed data, and spectrum libraries are publicly available on MASSIVE via accession MSV000090635.
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影响因子：
48
作者：
Choi M;Carver J;Chiva C;Tzouros M;Huang T;Tsai TH;Pullman B;Bernhardt OM;Hüttenhain R;Teo GC;Perez-Riverol Y;Muntel J;Müller M;Goetze S;Pavlou M;Verschueren E;Wollscheid B;Nesvizhskii AI;Reiter L;Dunkley T;Sabidó E;Bandeira N;Vitek O
通讯作者：
Vitek O
影响因子：
4.4
作者：
Kassem S;van der Pan K;de Jager AL;Naber BAE;de Laat IF;Louis A;van Dongen JJM;Teodosio C;Díez P
通讯作者：
Díez P
影响因子：
16.6
作者：
Schoof EM;Furtwängler B;Üresin N;Rapin N;Savickas S;Gentil C;Lechman E;Keller UAD;Dick JE;Porse BT
通讯作者：
Porse BT
影响因子：
7.3
作者：
Kuboki, Yuko;Fischer, Catherine G.;Wood, Laura D.
通讯作者：
Wood, Laura D.
影响因子：
56.9
作者：
Zhang, L;Zhou, W;Kinzler, KW
通讯作者：
Kinzler, KW