Challenges of the Human Proteome Project: 10-Year Experience of the Russian Consortium.
J Proteome Res
; 18(12): 4206-4214, 2019 12 06.
Article
em En
| MEDLINE
| ID: mdl-31599598
ABSTRACT
This manuscript collects all the efforts of the Russian Consortium, bottlenecks revealed in the course of the C-HPP realization, and ways of their overcoming. One of the main bottlenecks in the C-HPP is the insufficient sensitivity of proteomic technologies, hampering the detection of low- and ultralow-copy number proteins forming the "dark part" of the human proteome. In the frame of MP-Challenge, to increase proteome coverage we suggest an experimental workflow based on a combination of shotgun technology and selected reaction monitoring with two-dimensional alkaline fractionation. Further, to detect proteins that cannot be identified by such technologies, nanotechnologies such as combined atomic force microscopy with molecular fishing and/or nanowire detection may be useful. These technologies provide a powerful tool for single molecule analysis, by analogy with nanopore sequencing during genome analysis. To systematically analyze the functional features of some proteins (CP50 Challenge), we created a mathematical model that predicts the number of proteins differing in amino acid sequence proteoforms. According to our data, we should expect about 100â¯000 different proteoforms in the liver tissue and a little more in the HepG2 cell line. The variety of proteins forming the whole human proteome significantly exceeds these results due to post-translational modifications (PTMs). As PTMs determine the functional specificity of the protein, we propose using a combination of gene-centric transcriptome-proteomic analysis with preliminary fractionation by two-dimensional electrophoresis to identify chemically modified proteoforms. Despite the complexity of the proposed solutions, such integrative approaches could be fruitful for MP50 and CP50 Challenges in the framework of the C-HPP.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteínas
/
Proteoma
/
Proteômica
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
Limite:
Humans
País como assunto:
Asia
/
Europa
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article