[Apoptosis-inducing ligand TRAIL can be recruited to lipid rafts].
OBJECTIVE
To investigate the relationship between tumor necrosis factor-related apoptosis-inducing ligand(TRAIL) and cell membrane microdomain lipid rafts.


METHODS
The expression of TRAIL on K562 cells was detected by indirect immunofluorescence staining and flow cytometry. The lipid rafts on K562 cells were detected with FITC-labeled cholera toxin B subunit (CTx-FITC) which bound to the GM1 glycosphingolipid in lipid rafts. The lipid rafts were then cross-linked with an anti-FITC mAb after binding with CTx-FITC. The localization of TRAIL on K562 cells was analyzed with rabbit anti-TRAIL antibody, Cy3-labeled goat-anti-rabbit IgG (Cy3-labeled GAR) and laser scanning confocal microscope.


RESULTS
The cross-linking of lipid rafts appeared at 20 minutes, reached peak at 30 minutes, and weakened around 40 minutes. After the cells were labeled with CTx-FITC and anti-FITC mAb TRAIL was aggregated in the cross-linked lipid rafts.


CONCLUSIONS
The anti-FITC mAb could be used for cross-linking of lipid rafts. TRAIL can be recruited to lipid rafts.
Jia
Wei
W
Department of Immunology, Fourth Military Medical University, Xi'an 710032, China.
Zhuang
Ran
R
Li
Qi
Q
Cao
Yun-Xin
YX
Liu
Xue-Song
XS
Song
Chao-Jun
CJ
Jin
Bo-Quan
BQ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D017209:Apoptosis; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D020014:K562 Cells; D021962:Membrane Microdomains / Q000378:metabolism*; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D053221:TNF-Related Apoptosis-Inducing Ligand; D016650:Fluorescein-5-isothiocyanate