Structural characterization of lacticin 3147, a two-peptide lantibiotic with synergistic activity.
Lantibiotics are antibacterial peptides isolated from bacterial sources that exhibit activity toward Gram-positive organisms and are usually several orders of magnitude more potent than traditional antibiotics such as penicillin. They contain a number of unique structural features including dehydro amino acid and lanthionine (thioether) residues. Introduced following ribosomal translation of the parent peptide, these moieties render conventional methods of peptide analysis ineffective. We report herein a new method using nickel boride (Ni(2)B), in the presence of deuterium gas, to reduce dehydro side chains and reductively desulfurize lanthionine bridges found in lantibiotics. Using this approach, it is possible to identify and distinguish the original locations of dehydro side chains and lanthionine bridges by traditional peptide sequencing (Edman degradation) followed by mass spectrometry. The strategy was initially verified using nisin A, a structurally well characterized lantibiotic, and subsequently extended to the novel two-component lantibiotic, lacticin 3147, produced by Lactococcus lactis subspecies lactis DPC3147. The primary structures of both lacticin 3147 peptides were then fully assigned by use of multidimensional NMR spectroscopy, showing that lacticin 3147 A1 has a specific lanthionine bridging pattern which resembles the globular type-B lantibiotic mersacidin, whereas the A2 peptide is a member of the elongated type-A lantibiotic class. Also obtained by NMR were solution conformations of both lacticin 3147 peptides, indicating that A1 may adopt a conformation similar to that of mersacidin and that the A2 peptide adopts alpha-helical structure. These results are the first of their kind for a synergistic lantibiotic pair (only four such pairs have been reported to date).
Martin
Nathaniel I
NI
Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
Sprules
Tara
T
Carpenter
Michael R
MR
Cotter
Paul D
PD
Hill
Colin
C
Ross
R Paul
RP
Vederas
John C
JC
D000409:Alanine / Q000031:analogs & derivatives* / Q000737:chemistry; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001426:Bacterial Proteins / Q000737:chemistry*; D001430:Bacteriocins / Q000737:chemistry*; D001880:Boranes / Q000737:chemistry; D003903:Deuterium / Q000737:chemistry; D004357:Drug Synergism; D013294:Lactococcus lactis / Q000737:chemistry; D008969:Molecular Sequence Data; D009532:Nickel / Q000737:chemistry; D019906:Nuclear Magnetic Resonance, Biomolecular; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000737:chemistry; D011487:Protein Conformation; D012996:Solutions; D013329:Structure-Activity Relationship; D013440:Sulfides; D013455:Sulfur / Q000737:chemistry; D013816:Thermodynamics
D000596:Amino Acids; D001426:Bacterial Proteins; D001430:Bacteriocins; D001880:Boranes; D010446:Peptide Fragments; D012996:Solutions; D013440:Sulfides; C049893:nickel boride; C082508:lacticin 481; D013455:Sulfur; D009532:Nickel; D003903:Deuterium; C001520:lanthionine; D000409:Alanine