193 nm Ultraviolet Photodissociation for the Characterization of Singly Charged Proteoforms Generated by MALDI.
193 nm Ultraviolet Photodissociation for the Characterization of Singly Charged Proteoforms Generated by MALDI.
复制标题
193 nm 紫外光解离用于表征 MALDI 生成的单电荷蛋白质形式。
DOI：
10.1021/jasms.2c00302
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发表时间：
2023
影响因子：
3.2
通讯作者：
Paša-Tolić,Ljiljana
中科院分区：
文献类型：
--
作者：
Zemaitis,KevinJ;Zhou,Mowei;Kew,William;Paša-Tolić,Ljiljana
MALDI成像允许直接从微生物，植物和哺乳动物样品的近细胞分析。尽管检测到数百种变形，但仅用完整的质量信息识别未知仍然是一个明显的挑战，即使具有高质量分辨率和质量精度。为此，许多补充方法已被用于创建精确的质量匹配的实验数据库，包括批量或空间分辨自下而上和/或自上而下的蛋白质组学。在此，我们描述了193 nm紫外光解（UVPD）在UHMR HF Orbitrap上用MALDI-UVPD裂解四极分离单电荷泛素（m/z8565）。该平台不仅可以对终端碎片进行高分辨率精确的质量测量，还可以对大型内部碎片进行高分辨率精确的质量测量。概述的工作流程证明了自顶向下分析分离MALDI蛋白离子的可行性，以及在MALDI成像应用中更全面表征蛋白质形态的潜力。
MALDI imaging allows for the near-cellular profiling of proteoforms directly from microbial, plant, and mammalian samples. Despite detecting hundreds of proteoforms, identification of unknowns with only intact mass information remains a distinct challenge, even with high mass resolving power and mass accuracy. To this end, many supplementary methods have been used to create experimental databases for accurate mass matching, including bulk or spatially resolved bottom-up and/or top-down proteomics. Herein, we describe the application of 193 nm ultraviolet photodissociation (UVPD) for fragmentation of quadrupole isolated singly charged ubiquitin (m/z8565) by MALDI-UVPD on a UHMR HF Orbitrap. This platform permitted the high-resolution accurate mass measurement of not just terminal fragments but also large internal fragments. The outlined workflow demonstrates the feasibility of top-down analyses of isolated MALDI protein ions and the potential toward more comprehensive characterization of proteoforms in MALDI imaging applications.