ELECTROPERMEABILIZATION MEDIATES A STABLE INSERTION OF GLYCOPHORIN-A WITH CHINESE-HAMSTER OVARY CELL-MEMBRANES

Electropulsation allowed us to incorporate glycophorin A, an integral membrane protein, into mammalian nucleated cell membranes (Chinese ham ster ovary cells). The induction of stable protein association is effe ctive only when the field intensity is higher than its threshold value , creating membrane permeabilization to small molecules. Under control led conditions, cell viability was only slightly altered by this treat ment. Pulse number and duration controlled both the number of modified cells and incorporated molecules. The phenomena was temperature depen dent. An average of 5x10(4) molecules/cell was bound. About 80% of cel ls in the pulsed population were observed to incorporate glycophorin. The protein incorporation was shown to be stable 48 h after electroass ociation. Electrically bound proteins were shared between the cells af ter each division. As enhanced binding is detected if glycophorin is a dded after the pulses, it is the long-lived alteration of the membrane mediated by the pulses which supports the association.