Native Mass Spectrometry Imaging of Proteins and Protein Complexes by Nano-DESI.

Native Mass Spectrometry Imaging of Proteins and Protein Complexes by Nano-DESI.
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DOI:
10.1021/acs.analchem.0c05277
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发表时间:
2021-03-16
影响因子:
7.4
通讯作者:
Cooper HJ
Cooper HJ
中科院分区:
化学1区
文献类型:
--
作者:
Hale OJ;Cooper HJ

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在此之前,我们已经展示了天然质谱学成像(Native MSI),其中保持在天然类折叠构象中的蛋白质的空间分布是通过液体萃取表面分析(LESA)来确定的。虽然LESA为原理证明提供了一个很好的试验台,但目前成像的空间分辨率主要受采样移液管尖端的物理尺寸的限制。在这里,我们报告了采用纳米喷雾-解吸电喷雾电离(Nano-DESI)进行原生MSI,与原生Lesa MSI相比,在分辨率上有了实质性的提高。此外,天然的纳米DESI可用于直接从组织中进行定位的自上而下的蛋白质组学分析。蛋白质,包括以前没有被天然MSI检测到的同源二聚体复合体,通过碰撞激活、高分辨率MS和质子转移电荷减少的组合被鉴定。
Previously, we have demonstrated native mass spectrometry imaging (native MSI) in which the spatial distribution of proteins maintained in their native-like, folded conformations was determined using liquid extraction surface analysis (LESA). While providing an excellent testbed for proof of principle, the spatial resolution of LESA is currently limited for imaging primarily by the physical size of the sampling pipette tip. Here, we report the adoption of nanospray-desorption electrospray ionization (nano-DESI) for native MSI, delivering substantial improvements in resolution versus native LESA MSI. In addition, native nano-DESI may be used for location-targeted top–down proteomics analysis directly from tissue. Proteins, including a homodimeric complex not previously detected by native MSI, were identified through a combination of collisional activation, high-resolution MS and proton transfer charge reduction.
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