Rapid alterations in the plasma membrane structure of macrophages stimulated with bacterial lipopeptides.
Synthetic lipopeptide analogues of the N-terminal region of bacterial lipoprotein are potent activators of macrophages. In a previous study we showed that within minutes after their addition to macrophage cultures, lipopeptides were found attached to the plasma membranes and within different compartments of the cells. Their rapid interaction with the plasma membrane is thought to occur via the insertion of their three fatty acids. We used the freeze-fracture technique to study the influence of lipopeptides on the architecture of plasma membranes. Fifteen to thirty seconds after addition of the lipopeptides, the freeze-fractured plasma membranes show a rapid decrease in the particle density. This effect is not due to a loss of proteins, but is caused by lateral diffusion of single particles, which subsequently aggregate. These alterations are transient, temperature-sensitive and disappear 20 min after stimulation. At 4 degrees C, no change is found in the architecture of the plasma membranes. Using electron energy loss spectroscopy (EELS), lipopeptides can neither be detected on the membrane nor within the cells when incubated at this temperature. Our findings suggest that membrane protein aggregation is involved in the rapid uptake of lipopeptides into macrophages after their interaction with the plasma membranes.
Uhl
B
B
Institut für Immunobiologie, Arbeitsgruppe Medizinische Physik und Elektronenmikroskopie, Universität Freiburg, Deutschland.
Speth
V
V
Wolf
B
B
Jung
G
G
Bessler
W G
WG
Hauschildt
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001854:Bone Marrow Cells; D002462:Cell Membrane / Q000187:drug effects / Q000648:ultrastructure*; D003603:Cytotoxins / Q000494:pharmacology*; D005614:Freeze Fracturing; D008074:Lipoproteins / Q000494:pharmacology*; D008213:Lymphocyte Activation / Q000502:physiology*; D008264:Macrophages / Q000187:drug effects* / Q000648:ultrastructure; D051379:Mice; D008807:Mice, Inbred BALB C; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D013057:Spectrum Analysis; D013696:Temperature
D003603:Cytotoxins; D008074:Lipoproteins; C061110:N-palmitoyl-5,6-dipalmitoylcysteinyl-alanyl-glycine