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Number of detected proteins as the function of the sensitivity of proteomic technology in human liver cells

View ORCID ProfileNikita Vavilov, Ekaterina Ilgisonis, Andrey Lisitsa, Elena Ponomarenko, Tatiana Farafonova, Olga Tikhonova, View ORCID ProfileVictor Zgoda, Alexander Archakov
doi: https://doi.org/10.1101/2021.11.24.469687
Nikita Vavilov
1Institute of Biomedical Chemistry, Moscow, Russia, Pogodinskaya 10, 119121
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Ekaterina Ilgisonis
1Institute of Biomedical Chemistry, Moscow, Russia, Pogodinskaya 10, 119121
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Andrey Lisitsa
1Institute of Biomedical Chemistry, Moscow, Russia, Pogodinskaya 10, 119121
2East China University of Technology, Nanchang City, Jiangxi, 330013
3East-Siberian Research and Education Center, Tyumen, Russia, 625003
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Elena Ponomarenko
1Institute of Biomedical Chemistry, Moscow, Russia, Pogodinskaya 10, 119121
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Tatiana Farafonova
1Institute of Biomedical Chemistry, Moscow, Russia, Pogodinskaya 10, 119121
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Olga Tikhonova
1Institute of Biomedical Chemistry, Moscow, Russia, Pogodinskaya 10, 119121
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Victor Zgoda
1Institute of Biomedical Chemistry, Moscow, Russia, Pogodinskaya 10, 119121
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Alexander Archakov
1Institute of Biomedical Chemistry, Moscow, Russia, Pogodinskaya 10, 119121
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  • For correspondence: a.i.archakov@gmail.com
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Abstract

The main goal of the Russian part of C-HPP is to detect and functionally annotate missing proteins (PE2-PE4) encoded by human chromosome 18. However, identifying such proteins in a complex biological mixture using mass spectrometry (MS)-based methods is difficult due to the insufficient sensitivity of proteomic analysis methods. In this study, we determined the proteomic technology sensitivity using a standard set of UPS1 proteins as an example. The results revealed that 100% of proteins in a mixture could only be identified at a concentration of at least 10−9 M. The decrease in concentration leads to protein losses associated with technology sensitivity, and no UPS1 protein is detected at a concentration of 10−13 M. Therefore, two-dimensional fractionation of samples was applied to improve sensitivity. The human liver tissue was examined by selected reaction monitoring and shotgun methods of MS analysis using one-dimensional and two-dimensional fractionation to identify the proteins encoded by human chromosome 18. A total of 134 proteins were identified. The overlap between proteomic and transcriptomic data in human liver tissue was ~50%. This weak convergence is due to the low sensitivity of proteomic technology compared to transcriptomic approaches. Data is available via ProteomeXchange with identifier PXD026997.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted November 25, 2021.
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Number of detected proteins as the function of the sensitivity of proteomic technology in human liver cells
Nikita Vavilov, Ekaterina Ilgisonis, Andrey Lisitsa, Elena Ponomarenko, Tatiana Farafonova, Olga Tikhonova, Victor Zgoda, Alexander Archakov
bioRxiv 2021.11.24.469687; doi: https://doi.org/10.1101/2021.11.24.469687
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Number of detected proteins as the function of the sensitivity of proteomic technology in human liver cells
Nikita Vavilov, Ekaterina Ilgisonis, Andrey Lisitsa, Elena Ponomarenko, Tatiana Farafonova, Olga Tikhonova, Victor Zgoda, Alexander Archakov
bioRxiv 2021.11.24.469687; doi: https://doi.org/10.1101/2021.11.24.469687

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