Permeabilization of mammalian cells to proteins: poliovirus 2A(pro) as a probe to analyze entry of proteins into cells

Novoa, I., Benavente, J., Cotten, M. and Carrasco, L. (1997) Permeabilization of mammalian cells to proteins: poliovirus 2A(pro) as a probe to analyze entry of proteins into cells. Experimental Cell Research, 232(1), pp. 186-190. (doi:10.1006/excr.1997.3533) (PMID:9141636)

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Abstract

Two hybrid protein molecules containing the poliovirus protease 2A (MBP-2Apro) (maltose-binding protein-2Aproand MBP-Pseudomonasexotoxin A-2Apro) have been constructed and purified. Both hybrid proteins efficiently cleave the translation initiation factor eIF-4G when they are co-internalized into cells with adenovirus particles. Almost no intact eIF-4G can be detected in cells incubated with these proteins following this method. Reovirus infectious subviral particles also promote the delivery of MBP-2Aprointo cells, although less efficiently than adenovirus particles. None of the other methods employed to permeabilize cells to MBP-2Aproachieves the degree of eIF-4G cleavage observed with adenovirus particles. By comparison about 30% of cells electroporated with MBP-2Aprostill contain intact eIF-4G. More drastic electroporation conditions lead to a significant decrease of cell survival. Osmotic lysis of pinocytic vesicles resulted in 30% of the eIF-4G being cleaved in cells treated in suspension. Delivery of MBP-2Aproby pH-sensitive liposomes leads to poor hydrolysis of eIF-4G. Taken together our results indicate that permeabilization of cells with adenovirus particles is the most efficient method for introducing MBP-2Aprointo cells.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Cotten, Professor Matthew
Authors: Novoa, I., Benavente, J., Cotten, M., and Carrasco, L.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Experimental Cell Research
Publisher:Elsevier
ISSN:0014-4827
ISSN (Online):1090-2422

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