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  Broad range of missense error frequencies in cellular proteins.

Garofalo, R., Wohlgemuth, I., Pearson, M., Lenz, C., Urlaub, H., & Rodnina, M. V. (2019). Broad range of missense error frequencies in cellular proteins. Nucleic Acids Research, 47(6), 2932-2945. doi:10.1093/nar/gky1319.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0002-CA94-3 Version Permalink: http://hdl.handle.net/21.11116/0000-0003-B559-D
Genre: Journal Article

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 Creators:
Garofalo, R.1, Author              
Wohlgemuth, I.1, Author              
Pearson, M.1, Author              
Lenz, C.2, Author              
Urlaub, H.2, Author              
Rodnina, M. V.1, Author              
Affiliations:
1Department of Physical Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578598              
2Research Group of Bioanalytical Mass Spectrometry, MPI for Biophysical Chemistry, Max Planck Society, ou_578613              

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 Abstract: ssessment of the fidelity of gene expression is crucial to understand cell homeostasis. Here we present a highly sensitive method for the systematic Quantification of Rare Amino acid Substitutions (QRAS) using absolute quantification by targeted mass spectrometry after chromatographic enrichment of peptides with missense amino acid substitutions. By analyzing incorporation of near- and non-cognate amino acids in a model protein EF-Tu, we show that most of missense errors are too rare to detect by conventional methods, such as DDA, and are estimated to be between <10-7-10-5 by QRAS. We also observe error hotspots of up to 10-3 for some types of mismatches, including the G-U mismatch. The error frequency depends on the expression level of EF-Tu and, surprisingly, the amino acid position in the protein. QRAS is not restricted to any particular miscoding event, organism, strain or model protein and is a reliable tool to analyze very rare proteogenomic events.

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Language(s): eng - English
 Dates: 2019-01-152019-04-08
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1093/nar/gky1319
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Title: Nucleic Acids Research
Source Genre: Journal
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Pages: - Volume / Issue: 47 (6) Sequence Number: - Start / End Page: 2932 - 2945 Identifier: -