Disaggregation of endotoxin by isolated rat liver plasma membranes and lack of product binding.
The interaction of bacterial endotoxin with isolated rat liver cell membranes was studied to determine the extent and nature of any binding. Serratia marcescens endotoxin was incubated with isolated rat liver plasma membranes for varying periods of time at 37 C, and this mixture was then centrifuged through a discontinous sucrose density gradient. The membranes banded primarily at the 35 to 45% sucrose interface, whether or not they had been incubated with endotoxin. The endotoxin distributed itself throughout the gradient except when incubated with membranes, in which case it failed to sediment. This membrane-induced alteration in sedimentation could be prevented by heat inactivation of the membranes, and was found to be pH, time, temperature, and concentration dependent. There was neither associated degradation of the endotoxin, as measured by molecular sieve chromatography, nor loss in toxicity, as determined in lead-sensitized rats. These observations are consistent with an enzymatic disaggregation of the endotoxin by membranes and could represent a step in the uptake of the endotoxin by the reticuloendothelial system. No significant binding of this disaggregated endotoxin to the membranes could be detected, either after gradient separation or after repeated washing. This finding strongly suggests that, at least in cells that are active in the uptake of endotoxin, membrane-endotoxin interactions may be relatively transitory in nature, and that firm adherence to such membranes may not be a central feature of endotoxin toxicity.
D000818:Animals; D001665:Binding Sites; D002458:Cell Fractionation; D002462:Cell Membrane / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D002860:Chromium Radioisotopes; D004731:Endotoxins / Q000032:analysis / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008099:Liver / Q000378:metabolism / Q000648:ultrastructure*; D051381:Rats; D012706:Serratia marcescens
D002860:Chromium Radioisotopes; D004731:Endotoxins