Interaction of ostreolysin, a cytolytic protein from the edible mushroom Pleurotus ostreatus, with lipid membranes and modulation by lysophospholipids.
Ostreolysin is a 16-kDa cytolytic protein specifically expressed in primordia and fruiting bodies of the edible mushroom Pleurotus ostreatus. To understand its interaction with lipid membranes, we compared its effects on mammalian cells, on vesicles prepared with either pure lipids or total lipid extracts, and on dispersions of lysophospholipids or fatty acids. At nanomolar concentrations, the protein lysed human, bovine and sheep erythrocytes by a colloid-osmotic mechanism, compatible with the formation of pores of 4 nm diameter, and was cytotoxic to mammalian tumor cells. A search for lipid inhibitors of hemolysis revealed a strong effect of lysophospholipids and fatty acids, occurring below their critical micellar concentration. This effect was distinct from the capacity of ostreolysin to bind to and permeabilize lipid membranes. In fact, permeabilization of vesicles occurred only when they were prepared with lipids extracted from erythrocytes, and not with lipids extracted from P. ostreatus or pure lipid mixtures, even if lysophospholipids or fatty acids were included. Interaction with lipid vesicles, and their permeabilization, correlated with an increase in the intrinsic fluorescence and alpha-helical content of the protein, and with aggregation, which were not detected with lysophospholipids. It appears that either an unknown lipid acceptor or a specific lipid complex is required for binding, aggregation and pore formation. The inhibitory effect of lysophospholipids may reflect a regulatory role for these components on the physiological action of ostreolysin and related proteins during fruiting.
Sepcić
Kristina
K
CNR-ITC, Istituto di Biofisica - Sezione di Trento, Povo, Italy.
Berne
Sabina
S
Potrich
Cristina
C
Turk
Tom
T
Macek
Peter
P
Menestrina
Gianfranco
G
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000187:drug effects* / Q000378:metabolism; D005452:Fluoresceins / Q000378:metabolism; D005656:Fungal Proteins / Q000494:pharmacology; D006460:Hemolysin Proteins / Q000378:metabolism / Q000494:pharmacology*; D006461:Hemolysis / Q000502:physiology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D008246:Lysophospholipids / Q000378:metabolism*; D020076:Pleurotus / Q000737:chemistry*; D012756:Sheep; D017550:Spectroscopy, Fourier Transform Infrared; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D005452:Fluoresceins; D005656:Fungal Proteins; D006460:Hemolysin Proteins; D008051:Lipid Bilayers; D008246:Lysophospholipids; C461280:ostreolysin; C007740:fluorexon