Cyclic antimicrobial peptides based on Limulus anti-lipopolysaccharide factor for neutralization of lipopolysaccharide.
Bacterial endotoxin (lipopolysaccharide, LPS) is responsible for the septic shock syndrome. As potential therapeutic agents cyclic cationic antimicrobial peptides of different length, based on the Limulus anti-lipopolysaccharide factor (LALF), were synthesized, and their interaction with LPS was characterized physico-chemically and related to results in biological assays. All peptides inhibited the LPS-induced cytokine production in human mononuclear cells and the Limulus amebocyte lysate in a concentration-dependent way, with the peptide comprising the complete LPS-binding loop of the LALF (cLALF22) being the most effective. The peptides were neither cytotoxic nor hemolytic, except a slight effect of cLALF22. The peptides were able to displace Ca(2+) cations from a LPS monolayer, with cLALF22 being again most effective in accordance with results from isothermal titration calorimetry, in which saturation of binding was observed at an equimolar [cLALF22]:[LPS] ratio, and at a ratio 2-2.5 for the other peptides. For cLALF22, zeta (xi) potential experiments exhibited a complete compensation of the negative charges of LPS, whereas for the other peptides a residual negative potential of -20 to -40mV was found. X-ray diffraction experiments showed that the mixed unilamellar/cubic inverted aggregate structure of the lipid A part of LPS was converted into a multilamellar one. The gel to liquid crystalline phase transition of the acyl chains of LPS was changed upon cLALF22 binding, leading to a clear fluidization, which was not observed or only to a lesser degree for the other peptides. The affinity of the peptides for LPS led to a reduced binding of lipopolysaccharide-binding protein (LBP) to target membranes and hence to an inhibition of cytokine induction in human mononuclear cells.
Andrä
Jörg
J
Forschungszentrum Borstel, Leibniz-Zentrum für Medizin und Biowissenschaften, Biophysik, Parkallee 10, 23845 Borstel, Germany.
Lamata
Marta
M
Martinez de Tejada
Guillermo
G
Bartels
Rainer
R
Koch
Michel H J
MH
Brandenburg
Klaus
K
D023181:Antimicrobial Cationic Peptides; D060829:Arthropod Proteins; D006801:Humans; D007447:Invertebrate Hormones / Q000494:pharmacology*; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009500:Neutralization Tests / Q000379:methods; D010456:Peptides, Cyclic / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D010743:Phospholipids / Q000378:metabolism
D023181:Antimicrobial Cationic Peptides; D060829:Arthropod Proteins; D007447:Invertebrate Hormones; D008070:Lipopolysaccharides; D010456:Peptides, Cyclic; D010743:Phospholipids; C034705:antilipopolysaccharide factor (Limulus)