title: MS-READ: Quantitative measurement of amino acid incorporation.
abstract: Ribosomal protein synthesis results in the genetically programmed incorporation of amino acids into a growing polypeptide chain. Faithful amino acid incorporation that accurately reflects the genetic code is critical to the structure and function of proteins as well as overall proteome integrity. Errors in protein synthesis are generally detrimental to cellular processes yet emerging evidence suggest that proteome diversity generated through mistranslation may be beneficial under certain conditions. Cumulative translational error rates have been determined at the organismal level, however codon specific error rates and the spectrum of misincorporation errors from system to system remain largely unexplored. In particular, until recently technical challenges have limited the ability to detect and quantify comparatively rare amino acid misincorporation events, which occur orders of magnitude less frequently than canonical amino acid incorporation events. We now describe a technique for the quantitative analysis of amino acid incorporation that provides the sensitivity necessary to detect mistranslation events during translation of a single codon at frequencies as low as 1 in 10,000 for all 20 proteinogenic amino acids, as well as non-proteinogenic and modified amino acids. This article is part of a Special Issue entitled "Biochemistry of Synthetic Biology - Recent Developments" Guest Editor: Dr. Ilka Heinemann and Dr. Patrick O'Donoghue.
authors.affiliation: Department of Microbiology, The Ohio State University, 318 West 12th Avenue, Columbus, OH 43210, USA; Center for RNA Biology, The Ohio State University, 484 West 12th Avenue, Columbus, OH 43210, USA.
authors.forename: Kyle
authors.initials: K
authors.lastname: Mohler
authors.affiliation: Systems Biology Institute, Yale University, West Haven, CT 06516, USA; Department of Cellular & Molecular Physiology, Yale University School of Medicine, New Haven, CT 06520, USA.
authors.forename: Hans-Rudolf
authors.initials: HR
authors.lastname: Aerni
authors.affiliation: Systems Biology Institute, Yale University, West Haven, CT 06516, USA; Department of Cellular & Molecular Physiology, Yale University School of Medicine, New Haven, CT 06520, USA.
authors.forename: Brandon
authors.initials: B
authors.lastname: Gassaway
authors.affiliation: Department of Microbiology and Molecular Genetics, McGovern Medical School, University of Texas Health Science Center, Houston, TX 77030, USA.
authors.forename: Jiqiang
authors.initials: J
authors.lastname: Ling
authors.affiliation: Department of Microbiology, The Ohio State University, 318 West 12th Avenue, Columbus, OH 43210, USA; Center for RNA Biology, The Ohio State University, 484 West 12th Avenue, Columbus, OH 43210, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Ibba
authors.affiliation: Systems Biology Institute, Yale University, West Haven, CT 06516, USA; Department of Cellular & Molecular Physiology, Yale University School of Medicine, New Haven, CT 06520, USA. Electronic address: jesse.rinehart@yale.edu.
authors.forename: Jesse
authors.initials: J
authors.lastname: Rinehart
mesh_terms: D000596:Amino Acids / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D003062:Codon; D004926:Escherichia coli; D017930:Genes, Reporter; D005815:Genetic Code; D020125:Mutation, Missense / Q000502:physiology*; D014176:Protein Biosynthesis / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012441:Saccharomyces cerevisiae
chemical_list: D000596:Amino Acids; D003062:Codon; D011506:Proteins
keywords: Mass Spectrometry Reporter; Misincorporation; Mistranslation