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Cell Chem Biol. 2016 Mar 17;23(3):370-380. doi: 10.1016/j.chembiol.2015.11.017. Epub 2016 Feb 11.

Structure and tRNA Specificity of MibB, a Lantibiotic Dehydratase from Actinobacteria Involved in NAI-107 Biosynthesis.

Author information

1
Department of Biochemistry; University of Illinois at Urbana-Champaign, Roger Adams Laboratory, 600 S. Mathews Ave., Urbana IL, 61801, USA.
2
Department of Chemistry and Howard Hughes Medical Institute; University of Illinois at Urbana-Champaign, Roger Adams Laboratory, 600 S. Mathews Ave., Urbana IL, 61801,USA.
3
NAICONS Srl, Viale Ortles 22/4, 20139 Milan Italy.
4
Center for Biophysics and Computational Biology; University of Illinois at Urbana-Champaign, Roger Adams Laboratory, 600 S. Mathews Ave., Urbana IL, 61801, USA.
#
Contributed equally

Abstract

Class I lantibiotic dehydratases dehydrate selected Ser/Thr residues of a precursor peptide. Recent studies demonstrated the requirement of glutamyl-tRNA(Glu) for Ser/Thr activation by one of these enzymes (NisB) from the Firmicute Lactococcus lactis. However, the generality of glutamyl-tRNA(Glu) usage and the tRNA specificity of lantibiotic dehydratases have not been established. Here we report the 2.7-Å resolution crystal structure, along with the glutamyl-tRNA(Glu) utilization of MibB, a lantibiotic dehydratase from the Actinobacterium Microbispora sp. 107891 involved in the biosynthesis of the clinical candidate NAI-107. Biochemical assays revealed nucleotides A73 and U72 within the tRNA(Glu) acceptor stem to be important for MibB glutamyl-tRNA(Glu) usage. Using this knowledge, an expression system for the production of NAI-107 analogs in Escherichia coli was developed, overcoming the inability of MibB to utilize E. coli tRNA(Glu). Our work provides evidence for a common tRNA(Glu)-dependent dehydration mechanism, paving the way for the characterization of lantibiotics from various phyla.

PMID:
26877024
PMCID:
PMC4798866
DOI:
10.1016/j.chembiol.2015.11.017
[Indexed for MEDLINE]
Free PMC Article

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