Evidence for electro-induced membrane defects assessed by lateral mobility measurement of a GPi anchored protein.
Electrotransfer is a method by which molecules can be introduced into living cells via plasma membrane electropermeabilization. Here, we show that electropermeabilization affects the lateral mobility of Rae-1, a GPi anchored protein. Our results suggest that 10-20 % of the membrane surface is occupied by defects or pores and that these structures propagate rapidly (<1 min) over the cell surface. Electrotransfer of plasmid DNA (pDNA) also affects the lateral mobility of Rae-1. Furthermore, we clearly show that, once inserted into the plasma membrane, pDNA is completely immobile and excludes Rae-1; this indicates that the pDNA molecules are tightly packed together to form aggregates occupying at least the outer leaflet of the plasma membrane.
Institut de Pharmacologie et de Biologie Structurale, CNRS UMR 5089, 31077, Toulouse Cedex, France.
Jean Michel
JM
Escoffre
Marie
M
Hubert
Justin
J
Teissié
Marie Pierre
MP
Rols
Cyril
C
Favard
D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000378:metabolism; D018274:Electroporation; D029543:Nucleocytoplasmic Transport Proteins / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D016062:Porosity
D029543:Nucleocytoplasmic Transport Proteins; D004247:DNA