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Immunochemical Determination of Cellular Content of Translation Release Factor RF4 in Escherichia coli
Author(s) -
Andersen Lars Dyrskjot,
Moreno Juan Manuel Palacios,
Clark Brian F. C.,
Mortensen Kim Kusk,
SperlingPetersen Hans Uffe
Publication year - 1999
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/713803516
Subject(s) - escherichia coli , translation (biology) , chemistry , content (measure theory) , biochemistry , mathematics , gene , messenger rna , mathematical analysis
The biosynthesis of proteins in prokaryotes is terminated when a stop codon is present in the A‐site of the 70S ribosomal complex. Four different translation termination factors are known to participate in the termination process. Release factor RF1 and RF2 are responsible for the recognition of the stop codons, and RF3 is known to accelerate the overall termination process. Release factor RF4 is a protein involved in the release of the mRNA and tRNA from the ribosomal complex. Furthermore, RF4 is involved in the proofreading in the elongation step of protein biosynthesis. The cellular contents of RF1, RF2, and RF3 were determined earlier. Here we report the cellular content of RF4 in Escherichia coli to be ∼16 500 molecules per cell. The cells were grown in a rich medium and harvested in the beginning of the exponential growth phase. The quantifications were performed by using Western immunoblotting with radioactive iodinated streptavidin and biotinylated rabbit anti‐mouse immunoglobulins plus a highly specific monoclonal antibody against RF4 as first antibody.