Permeabilization Streptolysin

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... 

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. 

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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