Exposure of phosphatidylethanolamine on the surface of apoptotic cells.
In the early stages of apoptosis, phosphatidylserine (PS) is translocated from the inner side of the plasma membrane to the outer layer, which allows phagocytes to recognize and engulf the apoptotic cells. In this study we have analyzed the cell surface exposure of phosphatidylethanolamine (PE) in apoptotic CTLL-2 cells, a cytotoxic T cell line, using a tetracyclic polypeptide of 19 amino acids (Ro09-0198) which specifically recognizes the structure of PE and forms a tight equimolar complex with the phospholipid. Fluorescence microscopic analysis showed that the peptide, conjugated with fluorescence-labeled streptavidin (FL-SA-Ro), bound effectively to the cell surface of cells undergoing apoptosis in response to withdrawal of interleukin-2 from the culture media, but not to nonapoptotic cells. The binding of FL-SA-Ro to apoptotic cells was not uniform and the intense staining was observed on surface blebs of apoptotic cells. The FL-SA-Ro binding was inhibited specifically by liposomes containing PE, suggesting that PE is mainly exposed on the surface blebs of apoptotic cells. The specific binding of FL-SA-Ro to the apoptotic cells was also confirmed using a flourescence-activated cell sorter and the time-dependent cell surface exposure of PE correlated well with the exposure of PS, as detected by the binding of annexin V. This study provides the first direct evidence that PE as well as PS is exposed on the cell surface during the early stages of apoptosis, resulting in the total loss of asymmetric distribution of aminophospholipids in the plasma membrane bilayer.
Emoto
K
K
Department of Inflammation Research, Tokyo Metropolitan Institute of Medical Science (RINSHOKEN), Japan.
Toyama-Sorimachi
N
N
Karasuyama
H
H
Inoue
K
K
Umeda
M
M
D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects; D005434:Flow Cytometry; D007376:Interleukin-2 / Q000494:pharmacology; D008563:Membrane Lipids / Q000032:analysis*; D051379:Mice; D008810:Mice, Inbred C57BL; D010455:Peptides; D010456:Peptides, Cyclic; D010714:Phosphatidylethanolamines / Q000032:analysis*; D013602:T-Lymphocytes, Cytotoxic / Q000737:chemistry / Q000166:cytology* / Q000187:drug effects; D014407:Tumor Cells, Cultured / Q000187:drug effects
D000900:Anti-Bacterial Agents; D007376:Interleukin-2; D008563:Membrane Lipids; D010455:Peptides; D010456:Peptides, Cyclic; D010714:Phosphatidylethanolamines; C055996:lanthiopeptin