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Proteomic profiling of unnatural amino acid toxicity through differential expression of wild type vs. telluroproteins in Eschericia coli DL41(DE3)
Author(s) -
Brinkley Sarah Christine,
Boles Jeffrey O
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.522.2
Subject(s) - chemistry , auxotrophy , amino acid , methionine , proteomics , cytotoxicity , mass spectrometry , biochemistry , computational biology , escherichia coli , in vitro , biology , chromatography , gene
Three‐dimensional conformation of proteins can be solved with greater ease by utilizing unnatural amino acids (UAA), along with x‐ray crystallography. Because atoms naturally occurring in proteins do not have sufficient mass to interact with x‐ray radiation UAA, telluromethionine (TeMet), offers a solution to the “phasing problem”. Hijacking the molecular machinery of cells makes incorporation of TeMet easy, unfortunately, cytotoxicity of TeMet substantially reduces in vivo uptake. Differential expression of a methionine auxotroph, Eschericia coli DL41, can show differences between cells forced to use TeMet in the translation of proteins and cells grown in an authentic methionine environment. Protein profiling of these systems can reveal a molecular snapshot of E. coli DL41 at any given time. The condition of the cell, whether sick or healthy, young or old, can be viewed, quite literally, through proteomics. Detection with UPLC/qTOF MS enables the identification of changes in normal verses diseased states, allowing for the identification of proteins responsible for cell death. Toxic effects due to TeMet can then be reduced, thereby, providing a new tool for the elucidation of 3D structure.