Glycine to glutamic acid misincorporation observed in a recombinant protein expressed by Escherichia coli cells |
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Authors: | Huang Yunping O'Mara Brian Conover Matthew Ludwig Richard Fu Jinmei Tao Li Li Zheng Jian Rieble Siegfried Grace Michael J Russell Reb J |
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Institution: | Department of Biologics Product & Process Development, Bristol-Myers Squibb Technical Operations, Princeton, New Jersey 08543, USA. yunping.huang@bms.com |
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Abstract: | A novel amino acid misincorporation, in which the intended glycine (Gly) residues were replaced by a glutamic acid (Glu), was observed in a recombinant protein expressed by Escherichia coli. The misincorporation was identified by peptide mapping and liquid chromatography-tandem mass spectrometric analysis on proteolyzed peptides of the protein and verified using the corresponding synthetic peptides containing the misincorporated residues. Analysis of the distribution of the misincorporated residues and their codon usage shows strong correlation between this misincorporation and the use of rarely used codon within the E. coli expression system. Results in this study suggest that the usage of the rare codon GGA has resulted in a Glu for Gly misincorporation. |
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Keywords: | glycine to glutamic acid misincorporation recombinant Escherichia coli rare codon GGA |
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