HOPE-fixation of lung tissue allows retrospective proteome and phosphoproteome studies.
HOPE-fixation of lung tissue allows retrospective proteome and phosphoproteome studies.
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HOPE 固定肺组织可进行回顾性蛋白质组和磷酸蛋白质组研究
DOI：
10.1021/pr500096a
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发表时间：
2014
影响因子：
4.4
通讯作者：
L. Jänsch
中科院分区：
文献类型：
--
作者：
O. Shevchuk;N. Abidi;F. Klawonn;J. Wissing;M. Nimtz;C. KuglerM. Steinert;T. Goldmann;L. Jänsch
HEPES-谷氨酸缓冲液介导的有机溶剂保护效应(HOPE)固定已被引入作为临床标本福尔马林固定的替代方法。除了保存组织学的形态结构外，HOPE固定被证明与最近的RNA和DNA测序方法兼容。然而，到目前为止，HOPE固定材料用于质谱学检查蛋白质组的适宜性仍未确定。这一点特别令人感兴趣，因为蛋白质是药物研究的主要资源，可以为信号通路的活动状态提供有价值的见解。在这项研究中，我们从人肺组织中提取蛋白质，并通过蛋白质组和磷酸蛋白质组分析对HOPE处理和速冻组织进行了比较。来自精确质谱学的高置信度数据使2603个蛋白质和3036个磷酸化位点得以鉴定。HOPE固定并不妨碍蛋白质的代表性提取，研究它们的生化性质，包括亚细胞定位和细胞过程，发现固定类型没有引起偏见。总而言之，HOPE肺样本的蛋白质组和磷酸蛋白质组数据与快速冷冻组织获得的结果定性相同。因此，HOPE处理的组织在组织学和通过蛋白质组学对患者样本的回顾蛋白质组分析中都符合临床要求。
Hepes-glutamic acid buffer-mediated organic solvent protection effect (HOPE)-fixation has been introduced as an alternative to formalin fixation of clinical samples. Beyond preservation of morphological structures for histology, HOPE-fixation was demonstrated to be compatible with recent methods for RNA and DNA sequencing. However, the suitability of HOPE-fixed materials for the inspection of proteomes by mass spectrometry so far remained undefined. This is of particular interest, since proteins constitute a prime resource for drug research and can give valuable insights into the activity status of signaling pathways. In this study, we extracted proteins from human lung tissue and tested HOPE-treated and snap-frozen tissues comparatively by proteome and phosphoproteome analyses. High confident data from accurate mass spectrometry allowed the identification of 2603 proteins and 3036 phosphorylation sites. HOPE-fixation did not hinder the representative extraction of proteins, and investigating their biochemical properties, covered subcellular localizations, and cellular processes revealed no bias caused by the type of fixation. In conclusion, proteome as well as phosphoproteome data of HOPE lung samples were qualitatively equivalent to results obtained from snap-frozen tissues. Thus, HOPE-treated tissues match clinical demands in both histology and retrospective proteome analyses of patient samples by proteomics.
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影响因子：
2.8
作者：
Goldmann, T;Flohr, AM;Bullerdiek, J
通讯作者：
Bullerdiek, J
影响因子：
2.8
作者：
Goldmann, T;Wiedorn, KH;Vollmer, E
通讯作者：
Vollmer, E
肺部感染模型
影响因子：
1.2
作者：
P. Reinhold;M. Rosenbruch;D. Theegarten;K. Dalhoff;P. Dr;Dr. med. Petra Reinhold;Friedrich−Loeffler−Institut
通讯作者：
Friedrich−Loeffler−Institut
影响因子：
3.2
作者：
Marwitz, Sebastian;Abdullah, Mahdi;Goldmann, Torsten
通讯作者：
Goldmann, Torsten
影响因子：
4.6
作者：
Kothmaier, Hannelore;Rohrer, Daniela;Popper, Helmut H.
通讯作者：
Popper, Helmut H.