Amoebapores and NK-lysin, members of a class of structurally distinct antimicrobial and cytolytic peptides from protozoa and mammals: a comparative functional analysis.
Amoebapores, the pore-forming polypeptides of the protozoan parasite Entamoeba histolytica, and NK-lysin, an effector molecule of porcine NK (natural killer) and cytotoxic T cells, belong to the same protein family, the saposin-like proteins. As both types of protein are implicated in the killing of microbes in vivo, it appears that phylogenetically diverse organisms such as amoebae and mammals use similar effector molecules to fulfil a comparable task. However, structural features have led to the assumption that the proteins display their activities according to different modes of action. To address this question, we analysed the antibacterial, cytotoxic and pore-forming activities of these proteins in parallel and in comprehensive detail. Interestingly, the comparison of activities revealed significant differences. Whereas NK-lysin, recombinantly expressed, is efficient at a broad range of pH values, the amoebapores exhibited a pronounced pH dependence of all their activities, with markedly decreased activity at pH values above 6. Moreover, increasing salinity affects amoebapores more drastically than NK-lysin. All of the proteins compared were found to be potently active against Gram-positive bacteria, but only NK-lysin was equally efficient against Gram-negative bacteria. However, the amoebapores displayed five times higher pore-forming activity than NK-lysin, which is in accordance with the more hydrophobic character of the amoebapores compared with the essentially cationic NK-lysin.
Bruhn
Heike
H
Molecular Parasitology Group, Research Center for Infectious Diseases, Röntgenring 11, D-97070 Würzburg, Germany.
Riekens
Beate
B
Berninghausen
Otto
O
Leippe
Matthias
M
D000408:Alamethicin / Q000494:pharmacology; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001412:Bacillus subtilis / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004748:Entamoeba histolytica / Q000737:chemistry; D004926:Escherichia coli / Q000187:drug effects; D005452:Fluoresceins / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007473:Ion Channels / Q000302:isolation & purification / Q000494:pharmacology*; D019169:Jurkat Cells; D007963:Leukocytes, Mononuclear / Q000737:chemistry; D008081:Liposomes / Q000378:metabolism; D020033:Protein Isoforms / Q000302:isolation & purification / Q000494:pharmacology; D011510:Proteolipids / Q000302:isolation & purification / Q000494:pharmacology*; D015800:Protozoan Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D013552:Swine
D000900:Anti-Bacterial Agents; D005452:Fluoresceins; D007473:Ion Channels; D008081:Liposomes; C093238:NK-lysin; D020033:Protein Isoforms; D011510:Proteolipids; D015800:Protozoan Proteins; C041670:amoebapore proteins, protozoan; D000408:Alamethicin; C007740:fluorexon