Streptolysin Cell permeabilization

Permeabilized cells allow the study of intracellular processes in situ under conditions which are believed to be close to the physiological situation in intact cells. Permeabilization by bacterial pore-forming toxins, alpha-toxin and streptolysin O (SLO) is now a widely accepted approach in the functional analysis of intracellular organelles. 

Fohr KJ, Ahnert-Hilger G, Stecher B et al. (1991) GTP and Ca modulate the inositol 1,4,5,-trisphosphate-dependent Ca release in streptolysin O-permeabilized bovine adrenal chromaffin cells. J. Neurochem. 56 665—670. 

Stecher B, Ahnert-Hilger G, Weller U et at. (1992) Amylase release from streptolysin O-permeabilized pancreatic acinar cells effects of Ca GTPyS, cAMP, tetanus toxin and botulinum A toxin. Biochem. J. 283 899-904. 

Ahnert-Hilger G, Bader MF, BhakDai S et al. (1989b) Introduction of macromolecules into bovine adrenal medullary chromaffin cells and rat pheochromocy-toma cells (PC12) by permeabilization with streptolysin O inhibitory effect of tetanus toxin on catecholamine secretion. J. Neurochem. 52 1751 -8... 

The development of permeabilized cell systems has facilitated the reconstitution and biochemical characterization of a variety of intracellular protein trafficking steps. In this article I describe how coverslip-attached cells are permeabilized with streptolysin O (SLO) and incubated with exogenously added peroxisomal proteins. The protein import into peroxisomes is analyzed by a simple immunofluorescence technique. The procedure is described for CHO cells but it can be used for a variety of mammalian cells. Furthermore, the system can be applied to study the import of proteins into other subcellular organelles. 

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