Replacement of all arginine residues with canavanine in MazF-bs mRNA interferase changes its specificity.
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Ishida Y, Park JH, Mao L, Yamaguchi Y, Inouye M
Replacement of all arginine residues with canavanine in MazF-bs mRNA interferase changes its specificity.
J Biol Chem. 2013 Mar 15;288(11):7564-71. doi: 10.1074/jbc.M112.434969. Epub 2013 Feb 1.
- PubMed ID
- 23378533 [ View in PubMed]
- Abstract
Replacement of a specific amino acid residue in a protein with nonnatural analogues is highly challenging because of their cellular toxicity. We demonstrate for the first time the replacement of all arginine (Arg) residues in a protein with canavanine (Can), a toxic Arg analogue. All Arg residues in the 5-base specific (UACAU) mRNA interferase from Bacillus subtilis (MazF-bs(arg)) were replaced with Can by using the single-protein production system in Escherichia coli. The resulting MazF-bs(can) gained a 6-base recognition sequence, UACAUA, for RNA cleavage instead of the 5-base sequence, UACAU, for MazF-bs(arg). Mass spectrometry analysis confirmed that all Arg residues were replaced with Can. The present system offers a novel approach to create new functional proteins by replacing a specific amino acid in a protein with its analogues.