Methods for identifying modulators of apoptosis
Provided herein is a method for screening a modulator of apoptosis by contacting a cell comprising a FAS mediated apoptosis system, major vault protein and cytochrome b, with a candidate modulator, and measuring the level of apoptosis of the cell. The modulator of apoptosis is identified by a change in apoptosis in comparison to a control.
1. A method for screening a modulator of major vault protein comprising:
(a) providing a cell comprising major vault protein and a FAS-mediated apoptosis system, wherein the FAS-mediated apoptosis system comprises a FAS receptor and cytochrome b;
(b) providing a FAS ligand to the cell;
(c) contacting the cell with a candidate modulator compound;
(d) measuring the level of apoptosis of the cell; and
(e) comparing the level of apoptosis of the cell to a control;
wherein a modulator of major vault protein is identified by a change in apoptosis compared to the control.
2. The method of claim 1 , further comprising selecting the candidate modulator from a library of compounds.
3. The method of claim 2 , wherein the library of compounds is selected from the group consisting of a random peptide library, a natural products library, a cDNA library, a combinatorial library, an oligosaccharide library and a phage display library.
4. The method of claim 1 , further comprising the step of overexpressing the major vault protein.
5. The method of claim 1 , further comprising the step of expressing the major vault protein from heterologous DNA.
6. The method of claim 1 , further comprising the step of expressing the FAS-mediated apoptosis system from heterologous DNA.
7. The method of claim 6 , wherein the step of expressing the FAS-mediated apoptosis system comprises expressing the FAS receptor from a vector.
8. The method of claim 1 , further comprising the step of expressing the cytochrome b from a vector.
9. The method of claim 1 , wherein the method is performed in vitro.
10. The method of claim 1 , wherein the method is high throughput.
11. The method of claim 1 , wherein the FAS receptor ligand is selected from the group consisting of FAS receptor antibodies, FAS-L, FAS receptor specific ligands, and non-specific FAS receptor ligands.
12. The method of claim 1 , wherein the cell is non-mammalian.
13. The method of claim 1 , wherein the cell is mammalian.
14. The method of claim 13 , wherein the mammalian cell is selected from the group consisting of CHO cells and HeLa cells.
15. A method for screening a modulator of FAS-mediated apoptosis, comprising identifying a modulator of major vault protein according to the method of claim 1 , wherein a modulator of FAS-mediated apoptosis is identified as a modulator of major vault protein.