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)

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 > School of Infection & Immunity; College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research
• Journal Name: Experimental Cell Research
• Publisher: Elsevier
• ISSN: 0014-4827
• ISSN (Online): 1090-2422