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2013年更新:肝组织和HepG2细胞的18号染色体转录蛋白质组及血浆蛋白缺失后的靶向蛋白质组图谱

Chromosome 18 transcriptoproteome of liver tissue and HepG2 cells and targeted proteome mapping in depleted plasma: update 2013.

作者信息

Ponomarenko Elena A, Kopylov Arthur T, Lisitsa Andrey V, Radko Sergey P, Kiseleva Yana Yu, Kurbatov Leonid K, Ptitsyn Konstantin G, Tikhonova Olga V, Moisa Alexander A, Novikova Svetlana E, Poverennaya Ekaterina V, Ilgisonis Ekaterina V, Filimonov Alexey D, Bogolubova Nadezhda A, Averchuk Valentina V, Karalkin Pavel A, Vakhrushev Igor V, Yarygin Konstantin N, Moshkovskii Sergei A, Zgoda Victor G, Sokolov Alexey S, Mazur Alexander M, Prokhortchouck Egor B, Skryabin Konstantin G, Ilina Elena N, Kostrjukova Elena S, Alexeev Dmitry G, Tyakht Alexander V, Gorbachev Alexey Yu, Govorun Vadim M, Archakov Alexander I

机构信息

Orekhovich Institute of Biomedical Chemistry of the Russian Academy of Medical Sciences , 10 Pogodinskaya Street, Moscow 119121, Russia.

出版信息

J Proteome Res. 2014 Jan 3;13(1):183-90. doi: 10.1021/pr400883x. Epub 2013 Dec 13.

Abstract

We report the results obtained in 2012-2013 by the Russian Consortium for the Chromosome-centric Human Proteome Project (C-HPP). The main scope of this work was the transcriptome profiling of genes on human chromosome 18 (Chr 18), as well as their encoded proteome, from three types of biomaterials: liver tissue, the hepatocellular carcinoma-derived cell line HepG2, and blood plasma. The transcriptome profiling for liver tissue was independently performed using two RNaseq platforms (SOLiD and Illumina) and also by droplet digital PCR (ddPCR) and quantitative RT-PCR. The proteome profiling of Chr 18 was accomplished by quantitatively measuring protein copy numbers in the three types of biomaterial (the lowest protein concentration measured was 10(-13) M) using selected reaction monitoring (SRM). In total, protein copy numbers were estimated for 228 master proteins, including quantitative data on 164 proteins in plasma, 171 in the HepG2 cell line, and 186 in liver tissue. Most proteins were present in plasma at 10(8) copies/μL, while the median abundance was 10(4) and 10(5) protein copies per cell in HepG2 cells and liver tissue, respectively. In summary, for liver tissue and HepG2 cells a "transcriptoproteome" was produced that reflects the relationship between transcript and protein copy numbers of the genes on Chr 18. The quantitative data acquired by RNaseq, PCR, and SRM were uploaded into the "Update_2013" data set of our knowledgebase (www.kb18.ru) and investigated for linear correlations.

摘要

我们报告了俄罗斯染色体中心人类蛋白质组计划(C-HPP)联盟在2012 - 2013年取得的成果。这项工作的主要范围是对人类18号染色体(Chr 18)上的基因进行转录组分析,以及对来自三种生物材料(肝组织、肝癌衍生细胞系HepG2和血浆)的编码蛋白质组进行分析。肝组织的转录组分析使用了两个RNA测序平台(SOLiD和Illumina)独立进行,还通过液滴数字PCR(ddPCR)和定量逆转录PCR进行。Chr 18的蛋白质组分析是通过使用选择反应监测(SRM)定量测量三种生物材料中的蛋白质拷贝数来完成的(测量到的最低蛋白质浓度为10^(-13) M)。总共估计了228种主蛋白质的蛋白质拷贝数,包括血浆中164种蛋白质、HepG2细胞系中171种蛋白质以及肝组织中186种蛋白质的定量数据。大多数蛋白质在血浆中的含量为10^8拷贝/μL,而在HepG2细胞和肝组织中,中位数丰度分别为每细胞10^4和10^5个蛋白质拷贝。总之,对于肝组织和HepG2细胞,生成了一个“转录蛋白质组”,反映了Chr 18上基因的转录本和蛋白质拷贝数之间的关系。通过RNA测序、PCR和SRM获得的定量数据被上传到我们知识库(www.kb18.ru)的“Update_2013”数据集中,并对线性相关性进行了研究。

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