Divalent cation-sensitive pores formed by natural and synthetic melittin and by Triton X-100.
Leakage of ions and low-molecular-weight metabolites from Lettre cells is induced by synthetic melittin, as effectively as by melittin isolated from bee venom; in each case leakage is inhibited by Ca2+, Zn2+ or H+. Inhibition of leakage by divalent cations is reversible in that Lettre cells incubated with melittin (or with Triton X-100) in the presence of inhibitory amounts of Zn2+, when freed of Zn2+ by EGTA or by centrifugation, begin to leak (in Zn2(+)-sensitive manner). Electrorotation of Lettre cells is altered by melittin, compatible with membrane permeabilization; melittin plus Zn2+ does not alter electrorotation until Zn2+ (and unbound melittin) are removed. Melittin or Triton X-100 added to calcein-loaded liposomes induces leakage of calcein; divalent cations inhibit. Energy transfer between liposome-associated melittin and 2-, 7- or 12-(9-anthroyloxy)stearate (AS) is maximal with 12-AS; addition of Zn2+ has little effect. Circular dichroism spectra of melittin plus liposomes are unaffected by Zn2+. These results show that the formation of divalent cation-sensitive pores is not dependent on the presence of endogenous membrane proteins and that the action of divalent cations is not by displacement of melittin (or Triton) from the lipid bilayer.
Alder
G M
GM
Department of Cellular and Molecular Sciences, St. George's Hospital Medical School, London, U.K.
Arnold
W M
WM
Bashford
C L
CL
Drake
A F
AF
Pasternak
C A
CA
Zimmermann
U
U
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002413:Cations, Divalent / Q000494:pharmacology*; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D002498:Centrifugation; D002942:Circular Dichroism; D003902:Detergents; D004533:Egtazic Acid; D004735:Energy Transfer; D005456:Fluorescent Dyes; D007700:Kinetics; D008081:Liposomes / Q000378:metabolism; D008555:Melitten / Q000494:pharmacology*; D051379:Mice; D017830:Octoxynol; D011092:Polyethylene Glycols; D013229:Stearic Acids; D015032:Zinc / Q000494:pharmacology
D002413:Cations, Divalent; D003902:Detergents; D005456:Fluorescent Dyes; D008081:Liposomes; D013229:Stearic Acids; D008555:Melitten; D011092:Polyethylene Glycols; D004533:Egtazic Acid; C065154:9-(9-anthroyloxy)stearic acid; D017830:Octoxynol; D015032:Zinc; D002118:Calcium