Leakage of internal markers from erythrocytes and lipid vesicles induced by melittin, gramicidin S and alamethicin: a comparative study.
The membrane-disruptive capacities of melittin, derivatised melittins, alamethicin and gramicidin S have been compared for the human erythrocyte membrane and lipid vesicles of three different compositions (phosphatidylcholine, 85% phosphatidylcholine/15% phosphatidylserine, and a lipid analogue of the outer leaflet of the human erythrocyte membrane). The sensitivity to ionic strength, divalent metal ions and polylysine of release of fluorescent markers from liposomes and of haemoglobin from intact erythrocytes has been assayed. Acetyl melittin was found to he more effective than melittin in lysing phosphatidylcholine and phosphatidylcholine/phosphatidylserine vesicles, somewhat less effective in the lipid analogue and markedly less effective in lysing erythrocytes. Succinyl melittin was non-haemolytic, but was able to lyse lipid vesicles at a high concentration. Ca2+ inhibited melittin haemolysis at high ionic strength (150 mM NaCl), but produced a more complex response of stimulation followed by inhibition at low ionic strength. In lipid vesicles, Ca2+ either stimulated melittin lysis or was ineffective. Zn2+ exerted effects similar to Ca2+ with lipid vesicles at approx. 10-fold lower concentration except that a weak inhibition was observed for the erythrocyte membrane lipid analogue at high ionic strength. Polylysine strongly inhibited haemolysis by melittin at low ionic strength, but was ineffective or stimulatory in lipid vesicle lysis. High phosphate concentration also inhibited melittin haemolysis, but again no corresponding effect could he found in any of the lipid vesicle systems. These disparities between effects of melittin on erythrocytes and lipid vesicles support the proposal that melittin-protein interactions are of consequence to its haemolytic action. Similar experiments were performed with gramicidin S and alamethicin in order to compare their lytic properties with those of melittin. It was found that each lysin exhibited its own individual pattern of sensitivity to lipid composition, ionic strength and inhibition by cations. It thus appears likely that the detailed molecular interactions responsible for lysis are significantly different for each of these three agents.
Department of Chemistry and Biological Chemistry, University of Essex, Colchester, U.K.
S H
SH
Portlock
M J
MJ
Clague
R J
RJ
Cherry
D000408:Alamethicin / Q000494:pharmacology*; D000595:Amino Acid Sequence; D002118:Calcium / Q000494:pharmacology; D002413:Cations, Divalent; D004910:Erythrocyte Membrane / Q000187:drug effects* / Q000378:metabolism; D006096:Gramicidin / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D008081:Liposomes / Q000032:analysis / Q000378:metabolism*; D008555:Melitten / Q000031:analogs & derivatives / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009994:Osmolar Concentration; D010710:Phosphates / Q000494:pharmacology; D010713:Phosphatidylcholines / Q000032:analysis; D010718:Phosphatidylserines / Q000032:analysis; D011107:Polylysine / Q000494:pharmacology; D015032:Zinc / Q000494:pharmacology
D002413:Cations, Divalent; D008081:Liposomes; D010710:Phosphates; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D006096:Gramicidin; D008555:Melitten; D011107:Polylysine; D000408:Alamethicin; D015032:Zinc; D002118:Calcium