TY - JOUR
T1 - Structure and tRNA Specificity of MibB, a Lantibiotic Dehydratase from Actinobacteria Involved in NAI-107 Biosynthesis
AU - Ortega, Manuel A.
AU - Hao, Yue
AU - Walker, Mark C.
AU - Donadio, Stefano
AU - Sosio, Margherita
AU - Nair, Satish K.
AU - van der Donk, Wilfred
N1 - The authors thank Dr. Neha Garg and Dr. Bo Li for initial cloning of mib genes. We thank Dr. Tiit Lukk for data collection at C.H.E.S.S. (Ithaca, NY) and Keith Brister and colleagues for facilitating data collection at LS-CAT (Argonne National Labs, IL). We also thank Christopher J. Thibodeaux, Gabrielle N. Thibodeaux, and Madeline L\u00F3pez for helpful discussions, as well as members of the van der Donk, Martinis, and Silverman laboratories at UIUC. This work was supported by a grant from the National Institutes of Health ( R01 GM 058822 to W.A.v.d.D and R01 GM 079038 to S.K.N.), and by a grant from the European Commission (contract no. 245066 for FP7-KBBE-2009-3 ) to M.S. M.A.O. was supported by the NIGMS-NIH Chemistry-Biology Interface Training Grant ( 5T32-GM070421 ) and by the Ford Foundation . Y.H. was supported partially by a Lowell P. Hager fellowship from the Department of Biochemistry . A Bruker UltrafleXtreme MALDI TOF/TOF mass spectrometer was purchased in part with a grant from the NIH ( S10 RR027109 A ). The contents of this work are solely the responsibility of the authors and do not necessarily represent the official views of the NIH, Ford Foundation, or the European Commission. The authors declare a conflict of interest. S.D. and M.S. are employees of NAICONS.
PY - 2016/3/17
Y1 - 2016/3/17
N2 - Class I lantibiotic dehydratases dehydrate selected Ser/Thr residues of a precursor peptide. Recent studies demonstrated the requirement of glutamyl-tRNAGlu for Ser/Thr activation by one of these enzymes (NisB) from the Firmicute Lactococcus lactis. However, the generality of glutamyl-tRNAGlu 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-tRNAGlu 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 tRNAGlu acceptor stem to be important for MibB glutamyl-tRNAGlu 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 tRNAGlu. Our work provides evidence for a common tRNAGlu-dependent dehydration mechanism, paving the way for the characterization of lantibiotics from various phyla.
AB - Class I lantibiotic dehydratases dehydrate selected Ser/Thr residues of a precursor peptide. Recent studies demonstrated the requirement of glutamyl-tRNAGlu for Ser/Thr activation by one of these enzymes (NisB) from the Firmicute Lactococcus lactis. However, the generality of glutamyl-tRNAGlu 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-tRNAGlu 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 tRNAGlu acceptor stem to be important for MibB glutamyl-tRNAGlu 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 tRNAGlu. Our work provides evidence for a common tRNAGlu-dependent dehydration mechanism, paving the way for the characterization of lantibiotics from various phyla.
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U2 - 10.1016/j.chembiol.2015.11.017
DO - 10.1016/j.chembiol.2015.11.017
M3 - Article
C2 - 26877024
AN - SCOPUS:84965002440
SN - 2451-9456
VL - 23
SP - 370
EP - 380
JO - Cell chemical biology
JF - Cell chemical biology
IS - 3
ER -