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J Proteome Res. 2016 Nov 4;15(11):4030-4038. Epub 2016 Aug 29.

State of the Art of Chromosome 18-Centric HPP in 2016: Transcriptome and Proteome Profiling of Liver Tissue and HepG2 Cells.

Author information

1
Institute of Biomedical Chemistry , Pogodinskaya Street, 10, Moscow 119121, Russia.
2
Pirogov Russian National Research Medical University , Ostrovitianov Str. 1, Moscow 117997, Russia.
3
Center "Bioengineering" Russian Academy of Sciences , Prospect 60-let Oktyabrya, 7, Build.1, Moscow 119071, Russia.
4
Scientific Research Institute of Physical-Chemical Medicine , Malaya Pirogovskaya, 1a, Moscow 119435, Russia.

Abstract

A gene-centric approach was applied for a large-scale study of expression products of a single chromosome. Transcriptome profiling of liver tissue and HepG2 cell line was independently performed using two RNA-Seq platforms (SOLiD and Illumina) and also by Droplet Digital PCR (ddPCR) and quantitative RT-PCR. Proteome profiling was performed using shotgun LC-MS/MS as well as selected reaction monitoring with stable isotope-labeled standards (SRM/SIS) for liver tissue and HepG2 cells. On the basis of SRM/SIS measurements, protein copy numbers were estimated for the Chromosome 18 (Chr 18) encoded proteins in the selected types of biological material. These values were compared with expression levels of corresponding mRNA. As a result, we obtained information about 158 and 142 transcripts for HepG2 cell line and liver tissue, respectively. SRM/SIS measurements and shotgun LC-MS/MS allowed us to detect 91 Chr 18-encoded proteins in total, while an intersection between the HepG2 cell line and liver tissue proteomes was ∼66%. In total, there were 16 proteins specifically observed in HepG2 cell line, while 15 proteins were found solely in the liver tissue. Comparison between proteome and transcriptome revealed a poor correlation (R2 ≈ 0.1) between corresponding mRNA and protein expression levels. The SRM and shotgun data sets (obtained during 2015-2016) are available in PASSEL (PASS00697) and ProteomeExchange/PRIDE (PXD004407). All measurements were also uploaded into the in-house Chr 18 Knowledgebase at http://kb18.ru/protein/matrix/416126 .

KEYWORDS:

Human Proteome Project; RNA-Seq; human chromosome 18; proteome; quantitative PCR; selected reaction monitoring; transcriptome

PMID:
27527821
DOI:
10.1021/acs.jproteome.6b00380
[Indexed for MEDLINE]

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