Quantitative mouse brain proteomics using culture-derived isotope tags as internal standards

被引：171

作者：

Ishihama, Y ^{[1
]}

Sato, T ^{[1
]}

Tabata, T ^{[1
]}

Miyamoto, N ^{[1
]}

Sagane, K ^{[1
]}

Nagasu, T ^{[1
]}

Oda, Y ^{[1
]}

机构：

[1] Eisai & Co Ltd, Lab Seeds Finding Technol, Tsukuba, Ibaraki 3002635, Japan

来源：

NATURE BIOTECHNOLOGY | 2005年 / 23卷 / 05期

关键词：

D O I：

10.1038/nbt1086

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

An important challenge for proteomics is to be able to compare absolute protein levels across biological samples(1,2). Here we introduce an approach based on the use of culture-derived isotope tags (CDITs) for quantitative tissue proteome analysis. We cultured Neuro2A cells in a stable isotope-enriched medium and mixed them with mouse brain samples to serve as internal standards. Using CDITs, we identified and quantified a total of 1,000 proteins, 97 - 98% of which were expressed in both mouse whole brain and Neuro2A cells. CDITs also allow comprehensive and absolute protein quantification. Synthetic unlabeled peptides were used to quantify the corresponding proteins labeled with stable isotopes in Neuro2A cells, and the results were used to obtain the absolute amounts of 103 proteins in mouse whole brain. The expression levels correlated well with those in Neuro2A cells. Thus, the use of CDITs allows both relative and absolute quantitative proteome studies.

引用

页码：617 / 621

页数：5

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