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Replacement of all arginine residues with canavanine in MazF-bs mRNA interferase changes its specificity.

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.

SUBMITTER: Ishida Y

PROVIDER: S-EPMC3597797 | biostudies-literature | 2013 Mar

REPOSITORIES: biostudies-literature

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Replacement of all arginine residues with canavanine in MazF-bs mRNA interferase changes its specificity.

Ishida Yojiro Y Park Jung-Ho JH Mao Lili L Yamaguchi Yoshihiro Y Inouye Masayori M

The Journal of biological chemistry 20130201 11

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) ...[more]

PMID: 23378533

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