Streptolysins

Co-enzyme obtained from cultures of various strains of Streptococcus haemolyticus and capable of changing plasminogen into plasmin (complex enzyme mixture of streptokinase, streptodornase and streptolysin 0"). From fermentation liquors of hemolytic streptococci species Streptococcus haemolyticus), e. g. H 46 A. 

SLE Systemic lupus erythematosus SLe Sialyl Lewis X antigen SLO Streptolysin-O SLPI Secretory leucocyte protease inhibitor... 

When alamethicin is added to a ternary vesicle system comprising PDA, phospholipid, and lipopolysaccharide (LPS), the addition of polymyxin, an LPS-binding antibiotic, sensitizes the vesicles to alamethicin (Katz et al. 2003). Cholesterol-containing PDA liposomes have been used to colorimetrically detect streptolysin O, a cholesterol-dependent pore-forming toxin (Ma and Cheng 2005). 

Portnoy, et al. Capacity of listeriolysin O, streptolysin O and perfringolysin O to mediate growth of Bacillus subtilis within mammalian cells. Infect Immun 1992 60 2710-2717. 

Dekker LV, De Graan PN, Pijnappel P et al (1991) Noradrenaline release from streptolysin O-permeated rat cortical synaptosomes effects of calcium, phorbol esters, protein kinase inhibitors, and antibodies to the neuron-specific protein kinase C substrate B-50 (GAP-43). J Neurochem 56 1146-53... 

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... 

Binding studies have only been undertaken with alpha-toxin (Cassidy and Harshman, 1976 Hildebrand etai, 1991), aerolysin (Howard and Buckley, 1982), streptolysin O (Palmer etal., 1993) and perfringolysin... 

Certain cellular processes are specifically influenced by gradients of monovalent ions across the plasma membrane. It appears that stimulation of some such processes are actually counteracted by concomitant flux. Ca -dependent processes in turn may be abrogated when pores are large enough to permit rapid egress of cytoplasmic proteins. Therefore, it is useful to differentiate between three types of pores (a) those that are selectively permissive for monovalent ions (e.g. staphylococcal alpha-toxin) (b) those that are permissive for Ca and small molecules, but not for proteins (e.g. E. coli hemolysin) and (c) large pores that allow passage of macromolecules (e.g. streptolysin O). 

In the following, we will describe our current protocols for isolating staphylococcal alpha-toxin and streptolysin-O from bacterial culture supernatants. The protocol for isolating recombinant streptolysin-O from call has been published (Weller et al., 1996) and will not be... 

Storage of purified streptolysin O. The eluate from the hydrophobic chromatography column may be aliquoted and kept frozen at-70°C. Freeze drying of purified streptolysin O, without loss of activity (even of dilute samples), is possible if 0.1 % (w/v) bovine serum albumin is added. 

The advantages of PFTs are manifold. The effects of digitonin or saponin are normally difficult to predict, control or monitor. In contrast, the actions of PFTs are well characterized and easy to control. The toxins are easy to handle. The two agents currently used by most groups, alpha-toxin and streptolysin O, are stable and can be stored in lyophilized form tor years. No special procedures are required to... 

When very large pores are needed, streptolysin O is presently the most convenient toxin to apply. Streptolysin O pores are so large that cytoplasmic proteins rapidly egress from the cells however, intracellular organelles and the cytoskeleton remain, so that key cellular processes such as vesicle trafficking and movement of peptides amongst cellular compartments can be analyzed, e.g. with the use of antibodies. 

Alouf JE, Geoffroy C (1988) Production, purification and assay of streptolysin O. In Microbial Toxins Tools in Enzymology. Methods in Enzymology 165 (Harshman S, ed) pp 52-59, Academic Press, San Diego. 

Bhakdi S, Bayley H, Valeva A etal. (1996) Staphylococcal alpha-toxin, streptolysin O, and Escherichia coli hemolysin prototypes of pore forming bacterial cytoly-sins. In Arch Microbiol 165 73-79. 

Bhakdi S, Tranum-Jensen J, Sziegoleit A (1985) Mechanism of membrane damage by streptolysin-O. In Infect Immun 47 52—60. 

Palmer M, Valeva A, Kehoe M et al. (1995) Kinetics of streptolysin O assembly. In EurJ Biochem 231 388-395. 

Pinkney M, Kapur V, Smith J et al. (1995) Different forms of streptolysin O produced by Streptococcus pyogenes and by Escherichia coli expressing recombinant toxin cleavage by streptococcal cystein protease. In Infect Immun 63 2776-2779. 

Weller, U, Muller L, Messner M et al. (1996) Expression of active streptolysin-O in E. coli as MBP-SLO fusion protein. The N-terminal 70 amino acids are not required for hemolytic activity. In EurJ Biochem, 236 34—39. 

Application of Alpha-Toxin and Streptolysin O as Tools in Cell Biological Research... 

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. 

---