Interaction of eosinophil granule major basic protein with synthetic lipid bilayers: a mechanism for toxicity.
Eosinophil granule major basic protein (MBP) is a potent toxin for mammalian cells and helminths, but the mechanism of its toxicity is not known. Here we tested whether MBP toxicity is exerted through its effect on the lipid bilayer of its targets. Liposomes prepared from synthetic phospholipids were used as targets for MBP and their properties examined by fluorescence and circular dichroism (CD) spectroscopy. MBP caused a change in the temperature transition profiles of acidic liposomes (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidyl serine or an equimolar mixture of 1,2-dimyristoyl-sn-glycero-3-phosphocholine and 1,2-dimyristoyl-sn-glycero-3-phosphatidic acid) and induced their aggregation as shown by fluorescence resonance energy transfer experiments. The CD spectra and fluorescence characteristics of MBP itself were altered by its interaction with acidic lipids. Blue shifts in the emission maxima of the Trp, and of the dimethylaminonaphthyl moiety in acrylodan-labeled MBP, and a reduction in the effectiveness of quenching of Trp fluorescence by acrylamide were observed in the presence of acidic lipids. None of these effects were noted with zwitterionic lipids. This MBP: lipid bilayer interaction resulted in fusion and lysis of liposomes as indicated by the fluorescent indicator calcein. The results demonstrate that MBP associates with acidic lipids and that it disrupts, aggregates, fuses, and lyses liposomes prepared from such lipids. Such interaction might account for its wide range of toxicity.
Abu-Ghazaleh
R I
RI
Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota 55905.
Gleich
G J
GJ
Prendergast
F G
FG
D020106:Acrylamide; D000178:Acrylamides / Q000494:pharmacology; D001798:Blood Proteins / Q000502:physiology* / Q000633:toxicity; D002942:Circular Dichroism; D047091:Eosinophil Granule Proteins; D004804:Eosinophils / Q000502:physiology*; D005453:Fluorescence; D006801:Humans; D008051:Lipid Bilayers / Q000138:chemical synthesis / Q000378:metabolism*; D008081:Liposomes / Q000378:metabolism; D012260:Ribonucleases; D013696:Temperature
D000178:Acrylamides; D001798:Blood Proteins; D047091:Eosinophil Granule Proteins; D008051:Lipid Bilayers; D008081:Liposomes; D020106:Acrylamide; D012260:Ribonucleases