Interaction of immunologically-active lipopeptides with membranes.
Synthetic tripalmitoyl-S-glycerylcysteinyl (Pam3Cys) peptides are derived from the N-terminal part of bacterial lipoprotein and constitute polyclonal B-lymphocyte and macrophage activators. In order to elucidate the primary events of leukocyte activation, we investigated the biophysical interaction of lipopeptides containing spin labels or fluorescent markers with phosphatidylcholine vesicles or immune cells. Utilizing fluorescence microscopy and FACS analysis we found, that the surface of cells, after incubation with a fluorescein-labelled lipopeptide, was highly fluorescent. In addition, capping and patching was observed. Furthermore, fluorescence quenching experiments and electron paramagnetic resonance studies using vesicles incubated with lipopeptides suggested, that the peptide moiety and other more polar molecules linked to the lipo-amino acid are exposed to the hydrophilic compartment. These results show that in lipopeptide conjugates the Pam3Cys moiety acts as an efficient membrane anchor for molecules covalently coupled to it. The sequestering of the fatty-acid chains of the lipopeptide within the membrane is an early step of interaction, which might induce the uptake of the lipopeptide into the cell and the stimulation of immunocompetent cells.
Institut für Organische Chemie, Universität Tübingen, Germany.
J W
JW
Metzger
W H
WH
Sawyer
B
B
Wille
L
L
Biesert
W G
WG
Bessler
G
G
Jung
D000595:Amino Acid Sequence; D001402:B-Lymphocytes / Q000187:drug effects / Q000378:metabolism*; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000494:pharmacology*; D002352:Carrier Proteins / Q000737:chemistry / Q000494:pharmacology*; D002462:Cell Membrane / Q000378:metabolism*; D003545:Cysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D004578:Electron Spin Resonance Spectroscopy; D029968:Escherichia coli Proteins; D005227:Fatty Acids / Q000378:metabolism; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D008934:Mitogens / Q000494:pharmacology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism / Q000494:pharmacology*; D013010:Sonication; D014407:Tumor Cells, Cultured
D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D005227:Fatty Acids; D008934:Mitogens; D010455:Peptides; C051659:mitogenic bacterial lipoprotein, E coli; C038837:tripalmitoyl cysteine; D016650:Fluorescein-5-isothiocyanate; D003545:Cysteine