Structure and Dynamics of the Lantibiotic Mutacin 1140

Smith, Linda L.; Zachariah, Cherian; Thirumoorthy, Ramanan; Rocca, Jim; Novák, Jan; Hillman, Jeffrey D.; Edison, Arthur S.

doi:10.1021/bi034490u

Cited by 43 publications

(53 citation statements)

References 32 publications

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“…The crude extract was run on either a semiprep C 18 column (octyldecyl silane [C 18 ]; particle size, 5 m; 4.6 by 250 mm; Agilent Zorbax) or an analytical column, as previously reported (35). 2-Mercaptoethanol (BME) modification of mutacin 1140 peptide variants was done as previously reported (13,36). The lack of PTM cyclase activity would prevent lanthionine ring formation and result in the presence of cysteine thiols that can be selectively labeled.…”

Section: Methodsmentioning

confidence: 99%

“…Mutacin 1140 and nisin have structurally similar rings A and B (Fig. 1A), and both nisin and mutacin 1140 bind and abduct lipid II (8)(9)(10)(11)(12)(13). Mutacin 1140 has also been shown to be bactericidal to pathogenic bacteria, such as methicillin-resistant Staphylococcus aureus (MRSA), with no significant development of resistance (14).…”

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confidence: 99%

“…However, the core peptide of mutacin 1140 is 12 amino acids shorter than that of nisin. Mutacin 1140 has the same ability to bind to lipid II, but it does not form bacterial membrane pores like nisin does (12,13).…”

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confidence: 99%

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Biosynthesis and Transport of the Lantibiotic Mutacin 1140 Produced by Streptococcus mutans

et al. 2015

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Lantibiotics are ribosomally synthesized peptide antibiotics composed of an N-terminal leader peptide that is cleaved to yield the active antibacterial peptide. Significant advancements in molecular tools that promote the study of lantibiotic biosynthesis can be used in Streptococcus mutans. Herein, we further our understanding of leader peptide sequence and core peptide structural requirements for the biosynthesis and transport of the lantibiotic mutacin 1140. Our study on mutacin 1140 biosynthesis shows a dedicated secondary cleavage site within the leader peptide and the dependency of transport on core peptide posttranslational modifications (PTMs). The secondary cleavage site on the leader peptide is found at the ؊9 position, and secondary cleavage occurs before the core peptide is transported out of the cell. The coordinated cleavage at the ؊9 position was absent in a lanT deletion strain, suggesting that the core peptide interaction with the LanT transporter enables uniform cleavage at the ؊9 position. Following transport, the LanP protease was found to be tolerant to a wide variety of amino acid substitutions at the primary leader peptide cleavage site, with the exception of arginine at the ؊1 position. Several leader and core peptide mutations produced core peptide variants that had intermediate stages of PTM enzyme modifications, supporting the concept that PTM enzyme modifications, secondary cleavage, and transport are occurring in a highly coordinated fashion. IMPORTANCEMutacin 1140 belongs to the class I lantibiotic family of ribosomally synthesized and posttranslationally modified peptides (RiPPs). The biosynthesis of mutacin 1140 is a highly efficient process which does not lead to a discernible level of production of partially modified core peptide variants. The products isolated from an extensive mutagenesis study on the leader and core peptides of mutacin 1140 show that the posttranslational modifications (PTMs) on the core peptide occur under a highly coordinated dynamic process. PTMs are dictated by the distance of the core peptide modifiable residues from PTM enzyme active sites. The formation of lanthionine rings aids in the formation of successive PTMs, as was observed in a peptide variant lacking a Cterminal decarboxylation. Lantibiotics are a class of ribosomally synthesized peptide antibiotics produced by Gram-positive bacteria, such as Lactococcus lactis and Streptococcus mutans (1, 2). Lantibiotics are characterized by the presence of posttranslational modifications (PTMs), such as dehydrated residues and lanthionine rings. The modified residues 2,3-didehydroalanine (Dha) and 2,3-didehydrobutyrine (Dhb) are formed from dehydration of serines and threonines, respectively. The cyclization between a cysteine and either a Dha or a Dhb forms a lanthionine or a methyllanthionine ring, respectively. The biosynthetic gene cluster contains all the genes necessary to produce a lantibiotic. The lan operon for class I lantibiotics contains genes encoding the lantibiotic peptide (lanA); the...

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Section: Methodsmentioning

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Biosynthesis and Transport of the Lantibiotic Mutacin 1140 Produced by Streptococcus mutans

et al. 2015

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“…Work performed by Smith et al using the membrane mimetic solvent acetonitrile/water (80 :20) showed that mutacin 1140 has an overall horseshoe-like shape that is kinked at the "hinge region" between rings B and C (20). This shape is the result of a turn-like motif in the hinge region that folds the N-terminal rings A and B (the lipid II binding domain) toward the C-terminal overlapped rings C and D (20).…”

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confidence: 99%

“…This shape is the result of a turn-like motif in the hinge region that folds the N-terminal rings A and B (the lipid II binding domain) toward the C-terminal overlapped rings C and D (20). Interestingly, the flexibility of the hinge region is believed to be important for promoting lateral assembly of mutacin 1140 monomers.…”

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Site-Directed Mutations in the Lanthipeptide Mutacin 1140

Chen

Wilson-Stanford

Cromwell

et al. 2013

Appl Environ Microbiol

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The oral bacterium Streptococcus mutans, strain JH1140, produces the antibiotic mutacin 1140. Mutacin 1140 belongs to a group of antibiotics called lanthipeptides. More specifically, mutacin 1140 is related to the epidermin type A(I) lanthipeptides. Mutagenesis experiments of this group of lanthipeptides have been primarily restricted to the posttranslationally modified mesolanthionine and 3-methyllanthionine residues. Site-directed mutagenesis of the core peptide of mutacin 1140 was performed using the suicide vector pVA891. Substitutions of the N-terminal residue, the charged residue in the hinge region, and residues in ring A and intertwined rings C and D were investigated. A truncation and insertion of residues in ring A and intertwined rings C and D were also performed to determine whether or not they would alter the antimicrobial activity of the producing strain. Bioassays revealed that five of 14 mutants studied had improved antimicrobial activity against the indicator strain Micrococcus luteus ATCC 10240. MICs against Streptococcus mutans UA159, Streptococcus pneumoniae ATCC 27336, Staphylococcus aureus ATCC 25923, Clostridium difficile UK1, and Micrococcus luteus ATCC 10240 were determined for three mutacin 1140 variants that had the most significant increases in bioactivity in the M. luteus bioassay. This mutagenesis study of the epidermin group of lanthipeptides shows that antimicrobial activity can be significantly improved.

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