Sizing the radius of the pore formed in erythrocytes and lipid vesicles by the toxin sticholysin I from the sea anemone Stichodactyla helianthus.
The radius of the pore formed by sticholysin I and II (StI, StII) in erythrocytes and sticholysin I in lipid vesicles was investigated. The rate of colloid osmotic lysis of human erythrocytes, exposed to one of the toxins in the presence of sugars of different size, was measured. The relative permeability of each sugar was derived and the pore radius estimated with the Renkin equation. The radius was similar for sticholysin I and II and was independent of the reference sugar chosen and of the toxin concentration applied. It was also the same when erythrocytes were pretreated with different toxin doses in the presence of a polyethylene glycol (PEG) large enough to prevent lysis and thereafter transferred to solutions containing oligosaccharides of different size where they did lyse at different rates. The osmometric behavior of large unilamellar vesicles (LUV) was thereafter used to estimate the toxin lesion radius in a model system. LUV transferred to a hyperosmotic solution with a certain sugar immediately shrank and then re-swelled at a rate dependent on the bilayer permeability to water and sugar. When LUV were previously permeabilized with StI, only a fraction of them, namely those not carrying pores, continued to behave as osmometers. By increasing the size of the added sugar and approaching the pore radius, the fraction of osmometric LUV increased. Relative permeabilities were derived and used to estimate a channel radius around 1.2 nm, both for sugars and for PEGs. In conclusion the sticholysin pore has a constant size independent of toxin concentration and similar in natural and artificial membranes, suggesting it has a fixed predominant structure.
Tejuca
M
M
CNR-ITC, Centro di Fisica degli Stati Aggregati, I-38050 Povo (Trento), Italy.
Dalla Serra
M
M
Potrich
C
C
Alvarez
C
C
Menestrina
G
G
D000818:Animals; D003064:Cnidarian Venoms / Q000494:pharmacology*; D003102:Colloids; D004910:Erythrocyte Membrane / Q000187:drug effects*; D005453:Fluorescence; D006460:Hemolysin Proteins / Q000494:pharmacology*; D007425:Intracellular Membranes / Q000502:physiology; D007473:Ion Channels / Q000187:drug effects*; D007700:Kinetics; D008081:Liposomes; D008433:Mathematics; D008563:Membrane Lipids / Q000378:metabolism; D009844:Oligosaccharides / Q000502:physiology; D009930:Organic Chemicals; D009997:Osmotic Pressure; D011092:Polyethylene Glycols / Q000494:pharmacology; D012615:Sea Anemones; D013997:Time Factors
D003064:Cnidarian Venoms; D003102:Colloids; D006460:Hemolysin Proteins; D007473:Ion Channels; D008081:Liposomes; D008563:Membrane Lipids; D009844:Oligosaccharides; D009930:Organic Chemicals; C404794:sticholysin II; C121348:stycholysin I; D011092:Polyethylene Glycols