Enhanced Spatial Mapping of Histone Proteoforms in Human Kidney Through MALDI-MSI by High-Field UHMR-Orbitrap Detection.
Enhanced Spatial Mapping of Histone Proteoforms in Human Kidney Through MALDI-MSI by High-Field UHMR-Orbitrap Detection.
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DOI：
10.1021/acs.analchem.2c01034
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发表时间：
2022-09-20
影响因子：
7.4
通讯作者：
Pasa-Tolic, Ljiljana
中科院分区：
文献类型：
--
作者：
Zemaitis, Kevin J.;Velickovic, Dusan;Kew, William;Fort, Kyle L.;Reinhardt-Szyba, Maria;Pamreddy, Annapurna;Ding, Yanli;Kaushik, Dharam;Sharma, Kumar;Makarov, Alexander A.;Zhou, Mowei;Pasa-Tolic, Ljiljana
核心组蛋白包括H2A、H2B、H3和H4，是真核细胞内细胞修复、转录和复制的关键调节剂，在疾病的发病机制和细胞对环境刺激的反应中起着至关重要的作用。传统的基于质谱（MS）的自下而上和自上而下的蛋白质组学允许对蛋白质和含有蛋白质形态的翻译后修饰（PTM）进行全面鉴定。然而，这些方法难以保持近细胞的空间分布，因为它们通常需要激光捕获显微解剖（LCM）和先进的样品制备技术。在此，我们将基质辅助激光解吸/电离（MALDI）源与Thermo Scientific Q Exactive HF Orbitrap MS（升级了超高质量范围（UHMR）板）耦合在一起，首次演示了使用该台式质谱仪直接从组织中测量高达16.5 kDa的互补高分辨率精确质量（HR/AM）。该平台在整个检测质量范围内实现了同位素分辨率，提供了具有低ppm质量误差的可靠的异形分配，并且比其他傅立叶变换质量分析仪的占空比大大增加。在近细胞空间分辨率的人体肾脏切片上证实了核心组蛋白的蛋白质形态定位，在健康活检和肾细胞癌（RCC）包含肾切除术的切片中发现了组蛋白和其他蛋白质形态的几个关键分布。使用MALDI-MS成像（MSI）进行蛋白质形态定位展示了高通量准确鉴定蛋白质形态的几个步骤，并为在大质量范围内绘制整个组织切片的生物分子分布提供了一种新工具。
Core histones including H2A, H2B, H3, and H4 are key modulators of cellular repair, transcription, and replication within eukaryotic cells, playing vital roles in the pathogenesis of disease and cellular responses to environmental stimuli. Traditional mass spectrometry (MS)-based bottom-up and top-down proteomics allows for the comprehensive identification of proteins and of post-translational modification (PTM) harboring proteoforms. However, these methodologies have difficulties preserving near-cellular spatial distributions because they typically require laser capture microdissection (LCM) and advanced sample preparation techniques. Herein, we coupled a matrix-assisted laser desorption/ionization (MALDI) source with a Thermo Scientific Q Exactive HF Orbitrap MS upgraded with ultrahigh mass range (UHMR) boards for the first demonstration of complementary high-resolution accurate mass (HR/AM) measurements of proteoforms up to 16.5 kDa directly from tissues using this benchtop mass spectrometer. The platform achieved isotopic resolution throughout the detected mass range, providing confident assignments of proteoforms with low ppm mass error and a considerable increase in duty cycle over other Fourier transform mass analyzers. Proteoform mapping of core histones was demonstrated on sections of human kidney at near-cellular spatial resolution, with several key distributions of histone and other proteoforms noted within both healthy biopsy and a section from a renal cell carcinoma (RCC) containing nephrectomy. The use of MALDI-MS imaging (MSI) for proteoform mapping demonstrates several steps toward high-throughput accurate identification of proteoforms and provides a new tool for mapping biomolecule distributions throughout tissue sections in extended mass ranges.
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影响因子：
4
作者：
Kruse ARS;Spraggins JM
通讯作者：
Spraggins JM
影响因子：
7.4
作者：
Hale OJ;Cooper HJ
通讯作者：
Cooper HJ
影响因子：
7.4
作者：
Chen B;Brown KA;Lin Z;Ge Y
通讯作者：
Ge Y
激光捕获微分 - 毛细管区电泳串联质谱法（LCM-CZE-MS/MS）用于空间分辨自上而下的蛋白质组学：斑马鱼大脑的试验研究。
影响因子：
2.9
作者：
通讯作者：
--
影响因子：
20.1
作者：
Gillette TG;Hill JA
通讯作者：
Hill JA