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• W2318596598 startingPage "8165" @default.
• W2318596598 abstract "A mass spectrometry-based method was developed to measure amino acid substitutions directly in proteins down to a level of 0.001%. When applied to recombinant proteins expressed in Escherichia coli, monoclonal antibodies expressed in mammalian cells, and human serum albumin purified from three human subjects, the method revealed a large number of amino acid misincorporations at levels of 0.001–0.1%. The detected misincorporations were not random but involved a single-base difference between the codons of the corresponding amino acids. The most frequent base differences included a change from G to A, corresponding to a G(mRNA)/U(tRNA) base pair mismatch during translation. We concluded that under balanced nutrients, G(mRNA)/U(tRNA) mismatches at any of the three codon positions and certain additional wobble position mismatches (C/U and/or U/U) are the main causes of amino acid misincorporations. The hypothesis was tested experimentally by monitoring the levels of misincorporation at several amino acid sites encoded by different codons, when a protein with the same amino acid sequence was expressed in E. coli using 13 different DNA sequences. The observed levels of misincorporation were different for different codons and agreed with the predicted levels. Other less frequent misincorporations may occur due to G(DNA)/U(mRNA) mismatch during transcription, mRNA editing, U(mRNA)/G(tRNA) mismatch during translation, and tRNA mischarging." @default.
• W2318596598 created "2016-06-24" @default.
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• W2318596598 date "2013-10-31" @default.
• W2318596598 modified "2023-10-08" @default.
• W2318596598 title "G/U and Certain Wobble Position Mismatches as Possible Main Causes of Amino Acid Misincorporations" @default.
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• W2318596598 doi "https://doi.org/10.1021/bi401002c" @default.
• W2318596598 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/24128183" @default.
• W2318596598 hasPublicationYear "2013" @default.
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