title: 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.
authors: Department of Biochemistry, Robert Wood Johnson Medical School and Center for Advanced Biotechnology and Medicine, Piscataway, New Jersey 08854.
authors: Yojiro
authors: Y
authors: Ishida
authors: Department of Biochemistry, Robert Wood Johnson Medical School and Center for Advanced Biotechnology and Medicine, Piscataway, New Jersey 08854.
authors: Jung-Ho
authors: JH
authors: Park
authors: Department of Biochemistry, Robert Wood Johnson Medical School and Center for Advanced Biotechnology and Medicine, Piscataway, New Jersey 08854.
authors: Lili
authors: L
authors: Mao
authors: Department of Biochemistry, Robert Wood Johnson Medical School and Center for Advanced Biotechnology and Medicine, Piscataway, New Jersey 08854.
authors: Yoshihiro
authors: Y
authors: Yamaguchi
authors: Department of Biochemistry, Robert Wood Johnson Medical School and Center for Advanced Biotechnology and Medicine, Piscataway, New Jersey 08854. Electronic address: inouye@cabm.rutgers.edu.
authors: Masayori
authors: M
authors: Inouye
mesh_terms: D000595:Amino Acid Sequence; D001120:Arginine / Q000737:chemistry*; D001412:Bacillus subtilis / Q000201:enzymology*; D001483:Base Sequence; D001667:Binding, Competitive; D002172:Canavanine / Q000737:chemistry*; D002942:Circular Dichroism; D017931:DNA Primers / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000737:chemistry*; D004722:Endoribonucleases / Q000737:chemistry*; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry*; D007700:Kinetics; D013058:Mass Spectrometry / Q000379:methods; D008956:Models, Chemical; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010957:Plasmids / Q000378:metabolism; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D012333:RNA, Messenger / Q000737:chemistry* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D017931:DNA Primers; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C422955:MazF protein, E coli; D012333:RNA, Messenger; D002172:Canavanine; D001120:Arginine; D004722:Endoribonucleases
references: Chem Commun (Camb). 2011 Feb 14;47(6):1839-41
references: 21103555
references: Mol Syst Biol. 2006;2:2006.0008
references: 16738554
references: J Genet. 2010 Dec;89(4):527-37
references: 21273705
references: J Am Chem Soc. 2010 Mar 31;132(12):4086-8
references: 20218600
references: Nat Rev Microbiol. 2011 Sep 19;9(11):779-90
references: 21927020
references: J Biol Chem. 2005 Feb 4;280(5):3143-50
references: 15537630
references: J Mol Biol. 2003 May 16;328(5):1071-81
references: 12729742
references: Nat Commun. 2012 Jan 03;3:607
references: 22215082
references: Proteins. 2003 Nov 1;53(2):320-2
references: 14517982
references: J Biol Chem. 1989 Aug 15;264(23):13693-6
references: 2760038
references: Appl Environ Microbiol. 2010 Sep;76(18):6063-8
references: 20656860
references: J Biol Chem. 1989 Jun 15;264(17):9768-71
references: 2656711
references: PLoS Genet. 2011 Oct;7(10):e1002327
references: 22022286
references: FEBS Lett. 2011 Aug 4;585(15):2526-32
references: 21763692
references: Mol Cell. 2003 Apr;11(4):875-84
references: 12718874
references: J Biol Chem. 2003 Aug 22;278(34):32300-6
references: 12810711
references: Nat Protoc. 2006;1(6):2733-41
references: 17406529
references: Annu Rev Genet. 2011;45:61-79
references: 22060041
references: J Biol Chem. 1991 Aug 25;266(24):15684-7
references: 1874726
references: Mol Microbiol. 2007 Jul;65(2):373-85
references: 17590231
references: J Biol Chem. 2006 Dec 8;281(49):37559-65
references: 17020876
references: Science. 1989 Apr 14;244(4901):182-8
references: 2649980
references: J Am Chem Soc. 2010 Oct 27;132(42):14819-24
references: 20925334
references: Annu Rev Biochem. 2004;73:147-76
references: 15189139