Steroid structural requirements for interaction of ostreolysin, a lipid-raft binding cytolysin, with lipid monolayers and bilayers.
Ostreolysin, a cytolytic protein from the edible oyster mushroom (Pleurotus ostreatus), recognizes and binds specifically to membrane domains enriched in cholesterol and sphingomyelin (or saturated phosphatidylcholine). These events, leading to permeabilization of the membrane, suggest that a cholesterol-rich liquid-ordered membrane phase, which is characteristic of lipid rafts, could be its possible binding site. In this work, we present effects of ostreolysin on membranes containing various steroids. Binding and membrane permeabilizing activity of ostreolysin was studied using lipid mono- and bilayers composed of sphingomyelin combined, in a 1/1 molar ratio, with natural and synthetic steroids (cholesterol, ergosterol, beta-sitosterol, stigmasterol, lanosterol, 7-dehydrocholesterol, cholesteryl acetate, and 5-cholesten-3-one). Binding to membranes and lytic activity of the protein are both shown to be dependent on the intact sterol 3beta-OH group, and are decreased by introducing additional double bonds and methylation of the steroid skeleton or C17-isooctyl chain. The activity of ostreolysin mainly correlates with the ability of the steroids to promote formation of liquid-ordered membrane domains, and is the highest with cholesterol-containing membranes. Furthermore, increasing the cholesterol concentration enhanced ostreolysin binding in a highly cooperative manner, suggesting that the membrane lateral distribution and accessibility of the sterols are crucial for the activity of this new member of cholesterol-dependent cytolysins.
Rebolj
Katja
K
Department of Biology, Biotechnical Faculty, University of Ljubljana, Vecna pot 111, 1000 Ljubljana, Slovenia.
Ulrih
Natasa Poklar
NP
Macek
Peter
P
Sepcić
Kristina
K
D002152:Calorimetry, Differential Scanning; D002784:Cholesterol / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry; D006460:Hemolysin Proteins; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry*; D021962:Membrane Microdomains / Q000187:drug effects; D008567:Membranes, Artificial; D010539:Permeability; D013261:Sterols / Q000737:chemistry*; D013329:Structure-Activity Relationship
D005656:Fungal Proteins; D006460:Hemolysin Proteins; D008051:Lipid Bilayers; D008567:Membranes, Artificial; D013261:Sterols; C461280:ostreolysin; D002784:Cholesterol