Amphiphilic alpha-helices are important structural motifs in the alpha and beta peptides that constitute the bacteriocin lactococcin G--enhancement of helix formation upon alpha-beta interaction.
Lactococcin G (LcnG) is an antimicrobial substance (bacteriocin) consisting of two peptides, LcnG-alpha and LcnG-beta. The structures of intact LcnG-alpha and LcnG-beta as well as various fragments of these peptides were studied by circular dichroism (CD) under several conditions. All peptides had a non-structured conformation in aqueous solutions. In the presence of trifluoroethanol, dodecylphosphocholine micelles and (negatively charged) dioleoylglycerophosphoglycerol (Ole2GroPGro) liposomes, varying amounts of alpha-helical structure were induced. Comparisons of the various fragments showed that helicity was concentrated in those parts of LcnG-alpha and LcnG-beta that would become amphiphilic if an alpha-helical structure was adopted. In the presence of zwitterionic dioleoylglycerophosphocholine (Ole2GroPCho) liposomes, the peptides were much less (if at all) structured, suggesting that the excess of positive charge on the antimicrobial peptides needs to be compensated by an excess of negative charge on the membrane. The structuring of LcnG-alpha and LcnG-beta in the presence of Ole2GroPGro liposomes was considerably enhanced when both peptides were presented simultaneously to the membranes. Consecutive addition of the two peptides to Ole2GroPGro liposomes did not give this additional structuring, indicating that the individual LcnG-alpha and LcnG-beta peptides associate with the membrane in a virtually irreversible manner that makes them inaccessible for interaction with the complementary peptide. The results suggest that upon arrival at and interaction with the target membrane, LcnG-alpha and LcnG-beta form a complex that consists of approximately 50% amphiphilic alpha-helices.
Hauge
H H
HH
Department of Biochemistry, University of Oslo, Norway. hhhauge@krebs.uio.no
Nissen-Meyer
J
J
Nes
I F
IF
Eijsink
V G
VG
D001430:Bacteriocins / Q000737:chemistry*; D002942:Circular Dichroism; D046911:Macromolecular Substances; D008823:Micelles; D010715:Phosphatidylglycerols; D010767:Phosphorylcholine / Q000031:analogs & derivatives; D017433:Protein Structure, Secondary
D001430:Bacteriocins; D046911:Macromolecular Substances; D008823:Micelles; D010715:Phosphatidylglycerols; C110915:lactococcin G; D010767:Phosphorylcholine; C028810:dodecylphosphocholine; C051388:1,2-dioleoyl-sn-glycero-3-phosphoglycerol