F protein induced fusion of Sendai viral envelopes with mouse teratocarcinoma cells through LeX‐LeX interaction

FEBS Letters

Kumar, Mukesh¹  Sarkar, Debi P.¹ 

[1] Department of Biochemistry, University of Delhi South Campus, Benito Juarez Road, New Delhi - 110021, India

关键词: Sendai;    Virosome;    Lex/CD15;    Membrane fusion;    F;    fusion factor;    CHO;    Chinese hamster ovary;    DMEM;    Dulbecco's modified Eagle's medium;    DPBS;    Dulbecco's phosphate-buffered saline;    TX-100;    Triton X-100;    DTT;    dithiothreitol;    NBD-PE;    N-4-nitrobenzo-2-oxa-1;    3-diazole phosphatidylethanolamine;    FCS;    fetal calf serum;   

DOI  :  10.1016/0014-5793(96)00698-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The efficiency of membrane fusion between reconstituted Sendau viral envelopes containing only the fusion protein (F-virosomes) and the plasma membrane of mouse teratocarcinoma cells (F9) in culture was assessed using an assay based on the relief of self-quenching of a lipid probe incorporated in the F-virosomes. The potential of F-virosomes was also evaluated for a targeted cytosolic delivery of lysozyme to F9 cells. [¹²⁵I]Lysozyme entrapped into F-virosomes was taken to examine its fusion-mediated transfer to the F9 cells. Target specificity of the F-virosomes was confirmed by the interaction between the terminal Le^x moiety (Ga1β1 → 4(Fucα1 → 3)GlcNac) of F protein and the Le^x determinant on the membrane of F9 cells. Incubation of the loaded F-virosomes with cells led to fusion-mediated delivery, as inferred from the ability of cells to internalize lysozyme in the presence of azide (a potent inhibitor of endocytosis). These results suggest that carbohydrate-carbohydrate interaction is strong enough for target cell recognition followed by phospholipid bilayer melding induced by fusion glycoprotein of Sendai virus.

【 授权许可】

Unknown