Complete alanine scanning of the two-component lantibiotic lacticin 3147: generating a blueprint for rational drug design.
Lantibiotics are post-translationally modified antimicrobial peptides which are active at nanomolar concentrations. Some lantibiotics have been shown to function by targeting lipid II, the essential precursor of cell wall biosynthesis. Given that lantibiotics are ribosomally synthesized and amenable to site-directed mutagenesis, they have the potential to serve as biological templates for the production of novel peptides with improved functionalities. However, if a rational approach to novel lantibiotic design is to be adopted, an appreciation of the roles of each individual amino acid (and each domain) is required. To date no lantibiotic has been subjected to such rigorous analysis. To address this issue we have carried out complete scanning mutagenesis of each of the 59 amino acids in lacticin 3147, a two-component lantibiotic which acts through the synergistic activity of the peptides LtnA1 (30 amino acids) and LtnA2 (29 amino acids). All mutations were performed in situ in the native 60 kb plasmid, pMRC01. A number of mutations resulted in the elimination of detectable bioactivity and seem to represent an invariable core within these and related peptides. Significantly however, of the 59 amino acids, at least 36 can be changed without resulting in a complete loss of activity. Many of these are clustered to form variable domains within the peptides. The information generated in this study represents a blue-print that will be critical for the rational design of lantibiotic-based antimicrobial compounds.
Cotter
Paul D
PD
Department of Microbiology, University College Cork, Cork, Ireland.
Deegan
Lucy H
LH
Lawton
Elaine M
EM
Draper
Lorraine A
LA
O'Connor
Paula M
PM
Hill
Colin
C
Ross
R Paul
RP
D000409:Alanine / Q000031:analogs & derivatives / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D001430:Bacteriocins / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D017124:Conserved Sequence; D015195:Drug Design; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D013329:Structure-Activity Relationship; D013440:Sulfides / Q000378:metabolism
D001430:Bacteriocins; D013440:Sulfides; C082508:lacticin 481; C005080:beta-methyllanthionine; D000409:Alanine