Quantitative mouse brain proteomics using culture-derived isotope tags as internal standards.
An important challenge for proteomics is to be able to compare absolute protein levels across biological samples. 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.
Ishihama
Yasushi
Y
Sato
Toshitaka
T
Tabata
Tsuyoshi
T
Miyamoto
Norimasa
N
Sagane
Koji
K
Nagasu
Takeshi
T
Oda
Yoshiya
Y
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002460:Cell Line; D020869:Gene Expression Profiling / Q000379:methods* / Q000592:standards; D007553:Isotope Labeling / Q000379:methods* / Q000592:standards; D013058:Mass Spectrometry / Q000379:methods* / Q000592:standards; D051379:Mice; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism* / Q000592:standards; D040901:Proteomics / Q000379:methods; D012015:Reference Standards
D009419:Nerve Tissue Proteins; D020543:Proteome