Proteomic identification of immunoproteasome accumulation in formalin-fixed rodent spinal cords with experimental autoimmune encephalomyelitis.
Proteomic identification of immunoproteasome accumulation in formalin-fixed rodent spinal cords with experimental autoimmune encephalomyelitis.
复制标题
DOI：
10.1021/pr201043u
复制
发表时间：
2012-03-02
影响因子：
4.4
通讯作者：
Li, Hong
中科院分区：
文献类型：
--
作者：
Jain, Mohit Raja;Li, Qing;Liu, Tong;Rinaggio, Joseph;Ketkar, Amit;Tournier, Vincent;Madura, Kiran;Elkabes, Stella;Li, Hong
关键词：
临床相关的福尔马林固定和石蜡包埋（FFPE）组织尚未广泛用于神经蛋白质组学研究，因为许多蛋白质被认为是在组织保存过程中降解。蛋白质组学技术的最新改进，从完整蛋白质的2D凝胶分析到肽的“鸟枪法”定量以及绝对和相对定量（iTRAQ）方法的同量异序标签的使用，使得FFPE组织的分析成为可能。近年来，iTRAQ已成为高通量定量蛋白质组学分析的主要选择方法之一，该方法可在一个实验中同时比较多达8个样品。我们的目的是通过比较FFPE大鼠脊髓和冷冻组织中实验性自身免疫性脑脊髓炎（EAE）诱导的蛋白质组学变化，评估新鲜冷冻与FFPE神经组织iTRAQ分析的相对优点。FFPE组织中EAE诱导的蛋白质组变化与冷冻组织中发现的蛋白质组变化呈正相关，尽管蛋白质组覆盖率低约50%。随后对EAE脊髓中免疫蛋白酶体（IP）激活剂1富集的验证使我们评估了其他蛋白酶体和IP特异性蛋白。我们发现，许多IP特异性（而不是组成性）蛋白酶体蛋白在EAE大鼠脊髓中富集，EAE诱导的IP积累也发生在一个独立的小鼠EAE模型的脊髓中的残疾评分依赖性的方式。因此，我们的结论是，它是可行的，以产生有用的信息，从iTRAQ为基础的神经蛋白质组学分析存档FFPE组织研究神经疾病组织。
Clinically relevant formalin-fixed and paraffin-embedded (FFPE) tissues have not been widely used in neuroproteomic studies because many proteins are presumed to be degraded during tissue preservation. Recent improvements in proteomics technologies, from the 2D gel analysis of intact proteins to the “shotgun” quantification of peptides and the use of isobaric tags for absolute and relative quantification (iTRAQ) method has made the analysis of FFPE tissues possible. In recent years, iTRAQ has been one of the main methods of choice for high throughput quantitative proteomics analysis which enables simultaneous comparison of up to eight samples in one experiment. Our objective was to assess the relative merits of iTRAQ analysis of fresh frozen versus FFPE nervous tissues by comparing experimental autoimmune encephalomyelitis (EAE)-induced proteomic changes in FFPE rat spinal cords and frozen tissues. EAE-induced proteomic changes in FFPE tissues were positively correlated with those found in the frozen tissues, albeit with ~50% less proteome coverage. Subsequent validation of the enrichment of immunoproteasome (IP) activator 1 in EAE spinal cords led us to evaluate other proteasome and IP-specific proteins. We discovered that many IP-specific (as opposed to constitutive) proteasomal proteins were enriched in EAE rat spinal cords, and EAE-induced IP accumulation also occurred in the spinal cords of an independent mouse EAE model in a disability score-dependent manner. Therefore, we conclude that it is feasible to generate useful information from iTRAQ-based neuroproteomics analysis of archived FFPE tissues for studying neurological disease tissues.
登录
查看更多内容
影响因子：
3.3
作者：
Frausto, Ricardo F.;Crocker, Stephen J.;Whitton, J. Lindsay
通讯作者：
Whitton, J. Lindsay
影响因子：
3.3
作者：
Hosseini, H;André, P;Lotteau, V
通讯作者：
Lotteau, V
影响因子：
3.4
作者：
Ahram, M;Flaig, MJ;Emmert-Buck, MR
通讯作者：
Emmert-Buck, MR
影响因子：
7
作者：
Hood, BL;Darfler, MM;Krizman, DB
通讯作者：
Krizman, DB
影响因子：
3.7
作者：
Díaz-Hernández, M;Martín-Aparicio, E;Lucas, JJ
通讯作者：
Lucas, JJ