Proteomic characterization of post-mortem human brain tissue following ultracentrifugation-based subcellular fractionation.
Proteomic characterization of post-mortem human brain tissue following ultracentrifugation-based subcellular fractionation.
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DOI：
10.1093/braincomms/fcac103
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发表时间：
2022
影响因子：
4.8
通讯作者：
Carlyle, Becky C.
中科院分区：
文献类型：
--
作者：
Kandigian, Savannah E.;Ethier, Elizabeth C.;Kitchen, Robert R.;Lam, Tukiet T.;Arnold, Steven E.;Carlyle, Becky C.
人脑组织的蛋白质组学表征越来越多地用于识别各种神经系统疾病的潜在新型生物标志物和药物靶点。在全组织研究中，结果可能由最大和最丰富的细胞器或组织细胞类型组成的比例变化驱动。空间蛋白质组学方法通过考虑细胞定位的重要性，增强了我们对疾病机制和亚细胞水平上变化的信号通路的认识，细胞定位对蛋白质功能有着至关重要的影响。基于密度梯度的超离心方法允许亚细胞分级分离，并已用于细胞系、小鼠和人脑组织中，以定量特定富集细胞器（如突触前和突触后）中的数千种蛋白质。连续超离心方法允许分析来自同一生物样品的多个细胞器，并且据我们所知，以前还没有应用于冷冻的死后人脑组织。使用冷冻人体组织进行组织分离面临两个主要挑战，即死后间隔，在此期间蛋白质可能从其通常位置浸出到胞质溶胶中，以及冷冻，这导致膜破裂。尽管存在这些挑战，在这个概念验证研究中，我们发现大多数蛋白质可重复地分离成基于密度的粗离心级分，这些级分富含适当的细胞器标记物，并且可以在阿尔茨海默病个体和对照个体的组织之间观察到蛋白质定位的显著差异。空间蛋白质组学完成的冷冻死后脑组织的连续ultracentrapgation能够分配数以千计的脑蛋白质的大型膜细胞室，包括细胞核，线粒体和突触，和非膜胞质室。
Proteomic characterization of human brain tissue is increasingly utilized to identify potential novel biomarkers and drug targets for a variety of neurological diseases. In whole-tissue studies, results may be driven by changes in the proportion of the largest and most abundant organelles or tissue cell-type composition. Spatial proteomics approaches enhance our knowledge of disease mechanisms and changing signalling pathways at the subcellular level by taking into account the importance of cellular localization, which critically influences protein function. Density gradient-based ultracentrifugation methods allow for subcellular fractionation and have been utilized in cell lines, mouse and human brain tissue to quantify thousands of proteins in specific enriched organelles such as the pre- and post-synapse. Serial ultracentrifugation methods allow for the analysis of multiple cellular organelles from the same biological sample, and to our knowledge have not been previously applied to frozen post-mortem human brain tissue. The use of frozen human tissue for tissue fractionation faces two major challenges, the post-mortem interval, during which proteins may leach from their usual location into the cytosol, and freezing, which results in membrane breakdown. Despite these challenges, in this proof-of-concept study, we show that the majority of proteins segregate reproducibly into crude density-based centrifugation fractions, that the fractions are enriched for the appropriate organellar markers and that significant differences in protein localization can be observed between tissue from individuals with Alzheimer’s disease and control individuals. Spatial proteomics accomplished by serial ultracentrifugation of frozen post-mortem brain tissue is capable of assigning thousands of brain proteins to large membranous cellular compartments, including the nucleus, mitochondria and synapse, and to the non-membranous cytosolic compartment.
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影响因子：
16.2
作者：
Bai, Bing;Wang, Xusheng;Peng, Junmin
通讯作者：
Peng, Junmin
影响因子：
4
作者：
Hung, Mien-Chie;Link, Wolfgang
通讯作者：
Link, Wolfgang
基于质谱的方法用于绘制蛋白质亚细胞定位，揭示了小鼠原代神经元的空间蛋白质组。
影响因子：
8.8
作者：
Itzhak DN;Davies C;Tyanova S;Mishra A;Williamson J;Antrobus R;Cox J;Weekes MP;Borner GHH
通讯作者：
Borner GHH
DOI：
10.1111/j.2517-6161.1995.tb02031.x
发表时间：
1995-01-01
影响因子：
5.8
作者：
BENJAMINI, Y;HOCHBERG, Y
通讯作者：
HOCHBERG, Y
影响因子：
1.7
作者：
Bijur, GN;Jope, RS
通讯作者：
Jope, RS