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 hamster ovary cells). The induction of stable protein association is effective only when the field intensity is higher than its threshold value, creating membrane permeabilization to small molecules. Under controlled conditions, cell viability was only slightly altered by this treatment. Pulse number and duration controlled both the number of modified cells and incorporated molecules. The phenomena was temperature dependent. An average of 5 x 10(4) molecules/cell was bound. About 80% of cells in the pulsed population were observed to incorporate glycophorin. The protein incorporation was shown to be stable 48 h after electroassociation. Electrically bound proteins were shared between the cells after each division. As enhanced binding is detected if glycophorin is added after the pulses, it is the long-lived alteration of the membrane mediated by the pulses which supports the association.
el Ouagari
K
K
Département de Glycoconjugués et Biomembranes, UPR 8221 CNRS, Toulouse, France.
Benoist
H
H
Sixou
S
S
Teissie
J
J
D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism*; D002463:Cell Membrane Permeability; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D018274:Electroporation; D006021:Glycophorins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008856:Microscopy, Fluorescence; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013696:Temperature; D014357:Trypsin / Q000378:metabolism
D006021:Glycophorins; D012710:Serum Albumin, Bovine; D014357:Trypsin