Evidence for cooperativity of protein dissolution in Brij 58 permeabilized L929 cells.
Mouse L929 cells were exposed to the nonionic detergent Brij 58. As has been shown in some other cell types, protein leaked from Brij 58 exposed cells only after a lag phase. In the current study we have extended the observations of the kinetics of protein efflux using cultured L cells subjected to treatment with buffers containing Brij 58. The results show that while the cells become permeable essentially at first exposure to the detergent, proteins do not escape immediately. This lag in efflux is at least partly dependent on the concentration of detergent such that a greater lag is seen in cells exposed to the lowest concentrations of Brij. Data are presented that are most readily interpreted as protein leakage having occurred fairly rapidly from individual cells and that show that the time course of protein efflux results, to a large extent, from different sensitivities of individual cells to the detergent. The permeabilized suspension cells consist of only two types, whereas the conversion of cells from one type to the other occurs through the loss of protein to the permeabilization medium. Only two bands are seen in continuous density gradients and there is a conversion of the more dense type to the less dense with longer exposure to detergent. Moreover, the less dense cells contained about half of the protein per cell as the bottom banding cells, and the proteins of the more dense cells appear to be the sum of those released into the permeabilization medium plus those found in the less dense cells.
Ridsdale
J A
JA
University of California, Bodega Marine Laboratory, Bodega Bay 94923.
Clegg
J S
JS
D000818:Animals; D002021:Buffers; D002460:Cell Line; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002499:Centrifugation, Density Gradient; D002592:Cetomacrogol / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D005452:Fluoresceins / Q000378:metabolism; D007700:Kinetics; D007739:L Cells; D051379:Mice; D011506:Proteins / Q000378:metabolism*
D002021:Buffers; D005452:Fluoresceins; D011506:Proteins; C024098:6-carboxyfluorescein; D002592:Cetomacrogol