Interaction of antitumor alpha-lactalbumin-oleic acid complexes with artificial and natural membranes.
The specific complexes of human alpha-lactalbumin (alpha-LA) with oleic acid (OA), HAMLET and LA-OA-17 (OA-complexes), possess cytotoxic activity against tumor cells but the mechanism of their cell penetration remains unclear. To explore the molecular mechanisms underlying interaction of the OA-complexes with the cell membrane, their interactions with small unilamellar dipalmitoylphosphatidylcholine (DPPC) vesicles and electroexcitable plasma membrane of internodal native and perfused cells of the green alga Chara corallina have been studied. The fractionation (Sephadex G-200) of mixtures of the OA-complexes with the vesicles shows that OA-binding increases the affinity of alpha-LA to DPPC vesicles. Calcium association decreases protein affinity to the vesicles; the effect being less pronounced for LA-OA-17. The voltage clamp technique studies show that LA-OA-17, HAMLET, and their constituents produce different modifying effects on the plasmalemmal ionic channels of the Chara corallina cells. The irreversible binding of OA-complexes to the plasmalemma is accompanied by changes in the activation-inactivation kinetics of developing integral transmembrane currents, suppression of the Ca(2+) current and Ca(2+)-activated Cl(-) current, and by increase in the nonspecific K(+) leakage currents. The latter reflects development of nonselective permeability of the plasma membrane. The HAMLET-induced effects on the plasmalemmal currents are less pronounced and potentiated by LA-OA-17. The control experiments with OA and intact alpha-LA show their qualitatively different and much less pronounced effects on the transmembrane ionic currents. Thus, the modification of alpha-LA by OA results in an increase in the protein association with the model lipid bilayer and in drastic irreversible changes in permeability of several types of the plasmalemmal ionic channels.
Zherelova
Olga M
OM
Institute of Theoretical and Experimental Biophysics of the RAS, Pushchino, Moscow Region, Russia.
Kataev
Anatoly A
AA
Grishchenko
Valery M
VM
Knyazeva
Ekaterina L
EL
Permyakov
Sergei E
SE
Permyakov
Eugene A
EA
D015060:1,2-Dipalmitoylphosphatidylcholine; D002118:Calcium / Q000378:metabolism; D002458:Cell Fractionation; D002462:Cell Membrane / Q000378:metabolism*; D002463:Cell Membrane Permeability / Q000502:physiology*; D044784:Chara / Q000166:cytology*; D006801:Humans; D017136:Ion Transport; D007700:Kinetics; D007768:Lactalbumin / Q000378:metabolism*; D008567:Membranes, Artificial; D009829:Oleic Acids / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D022161:Transport Vesicles / Q000378:metabolism
C506422:HAMLET complex, human; D008567:Membranes, Artificial; D009829:Oleic Acids; D015060:1,2-Dipalmitoylphosphatidylcholine; D007768:Lactalbumin; D002118:Calcium