The long-term goal of this research proposal is to provide insights into the molecular basis for antibiotic biosynthesis in bacteria, which will serve as a framework for the design of novel antibiotics. Recent work in biosynthetic combinatorial chemistry technology has already established the utility of bacterial polyketide synthases and nonribosomal peptide synthases in generating natural and un-natural products with antibiotic capabilities. We seek to extend the research of biosynthetic production of therapeutics by providing a framework for the use of lantibiotics, a family of ribosomally synthesized but post-translationally modified peptides. The engineering of these lantibiotics is complicated by the fact that any modification that are generated in the peptide precursor may affect its function as a substrate for the post-translational modification enzymes, and compromise the effectiveness of this approach. In order to delineate the substrate requirements for the post-translation modification enzyme, we aim to determine the high-resolution crystal structures of many of the lantibiotic modification enzymes. The crystal structures of these enzymes will be used to identify and design site-specific variants. We will test our structure-based hypothesis by correlating the structure with functional data generated in the laboratory of our collaborator. Additionally, we will determine the crystal structure of the immunity component that protects the lantibiotic-producing bacteria from the action of its self-produced drug. We seek to understand why lantibiotics that are nearly identical in sequence utilize immunity components that share no homology and vary drastically in size. Finally, we will utilize the mechanistic and structural data generated in the previous components of the research plan to engineer the production of novel lantibiotics. In this last aim, we will seek to utilize the principles of biosynthetic combinatorial chemistry to answer the question """"""""Can novel lanthionine-bearing lantibiotics be designed from structure-based principles?""""""""

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM079038-05
Application #
7769481
Study Section
Synthetic and Biological Chemistry B Study Section (SBCB)
Program Officer
Jones, Warren
Project Start
2006-02-01
Project End
2011-01-31
Budget Start
2010-02-01
Budget End
2011-01-31
Support Year
5
Fiscal Year
2010
Total Cost
$194,846
Indirect Cost
Name
University of Illinois Urbana-Champaign
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
041544081
City
Champaign
State
IL
Country
United States
Zip Code
61820
Ortega, Manuel A; Cogan, Dillon P; Mukherjee, Subha et al. (2017) Two Flavoenzymes Catalyze the Post-Translational Generation of 5-Chlorotryptophan and 2-Aminovinyl-Cysteine during NAI-107 Biosynthesis. ACS Chem Biol 12:548-557
Repka, Lindsay M; Chekan, Jonathan R; Nair, Satish K et al. (2017) Mechanistic Understanding of Lanthipeptide Biosynthetic Enzymes. Chem Rev 117:5457-5520
Ortega, Manuel A; Hao, Yue; Walker, Mark C et al. (2016) Structure and tRNA Specificity of MibB, a Lantibiotic Dehydratase from Actinobacteria Involved in NAI-107 Biosynthesis. Cell Chem Biol 23:370-380
Dong, Shi-Hui; Tang, Weixin; Lukk, Tiit et al. (2015) The enterococcal cytolysin synthetase has an unanticipated lipid kinase fold. Elife 4:
Ortega, Manuel A; Hao, Yue; Zhang, Qi et al. (2015) Structure and mechanism of the tRNA-dependent lantibiotic dehydratase NisB. Nature 517:509-12
van der Donk, Wilfred A; Nair, Satish K (2014) Structure and mechanism of lanthipeptide biosynthetic enzymes. Curr Opin Struct Biol 29:58-66
Ortega, Manuel A; Velásquez, Juan E; Garg, Neha et al. (2014) Substrate specificity of the lanthipeptide peptidase ElxP and the oxidoreductase ElxO. ACS Chem Biol 9:1718-25
Garg, Neha; Tang, Weixin; Goto, Yuki et al. (2012) Lantibiotics from Geobacillus thermodenitrificans. Proc Natl Acad Sci U S A 109:5241-6
Li, Zhi; Rupasinghe, Sanjeewa G; Schuler, Mary A et al. (2011) Crystal structure of a phenol-coupling P450 monooxygenase involved in teicoplanin biosynthesis. Proteins 79:1728-38
Zou, Yaozhong; Nair, Satish K (2009) Molecular basis for the recognition of structurally distinct autoinducer mimics by the Pseudomonas aeruginosa LasR quorum-sensing signaling receptor. Chem Biol 16:961-70

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