Spatially-directed protein identification from tissue sections by top-down LC-MS/MS with electron transfer dissociation.
Spatially-directed protein identification from tissue sections by top-down LC-MS/MS with electron transfer dissociation.
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DOI：
10.1021/ac400832w
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发表时间：
2013-07-16
影响因子：
7.4
通讯作者：
Rose, Kristie L.
中科院分区：
文献类型：
--
作者：
Schey, Kevin L.;Anderson, David M.;Rose, Kristie L.
MALDI成像质谱（MALDI-IMS）已成为一个强大的工具，定位小分子和完整的蛋白质在各种各样的组织样本在正常和患病状态。MALDI-IMS中成像信号的识别仍然是分析中的瓶颈，并且限制了对组织标本的基础生物学的解释。在这项工作中，完整的蛋白质的空间定向组织微萃取，然后与电子转移解离（ETD）的LC-MS/MS被用来确定蛋白质从特定位置在三种组织类型：眼透镜，大脑和肾脏。检测限是这样的，1微升提取体积足以将蛋白质递送至LC-MS/MS仪器，其具有足够的灵敏度以在单个实验中检测50-100种蛋白质。此外，还鉴定了多种修饰的蛋白质;包括截短的透镜蛋白质，使用自下而上的方法难以将其分配到成像质量中。预期蛋白质分离和鉴定随着完整蛋白质分级分离/色谱法的进步和解释算法的进步而改进，从而导致来自不同组织位置的蛋白质组覆盖的深度增加。
MALDI imaging mass spectrometry (MALDI-IMS) has become a powerful tool for localizing both small molecules and intact proteins in a wide variety of tissue samples in both normal and diseased states. Identification of imaged signals in MALDI-IMS remains a bottleneck in the analysis and limits the interpretation of underlying biology of tissue specimens. In this work, spatially-directed tissue microextraction of intact proteins followed by LC-MS/MS with electron transfer dissociation (ETD) was used to identify proteins from specific locations in three tissue types; ocular lens, brain, and kidney. Detection limits were such that a 1 microliter extraction volume was sufficient to deliver proteins to the LC-MS/MS instrumentation with sufficient sensitivity to detect 50–100 proteins in a single experiment. Additionally, multiple modified proteins were identified; including truncated lens proteins that would be difficult to assign to an imaged mass using a bottom-up approach. Protein separation and identification are expected to improve with advances in intact protein fractionation/chromatography and advances in interpretation algorithms leading to increased depth of proteome coverage from distinct tissue locations.
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影响因子：
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Grey AC;Schey KL
通讯作者：
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影响因子：
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作者：
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DOI：
10.1016/s1044-0305(00)00223-3
发表时间：
2001-03-01
影响因子：
3.2
作者：
McLafferty, FW;Horn, DM;Carpenter, BK
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