Use of hepatitis B X-interacting protein (HBXIP) in modulation of apoptosis
Novel methods of regulating cellular apoptosis by affecting the interaction of hepatitis B X-interacting protein (HBXIP) with Survivin are described. More specifically, these novel methods of enhancing apoptosis of neoplastic cells comprises inhibiting interaction of hepatitis B X-interacting protein (HBXIP) with Survivin.
1. A method for identifying an effective agent that alters the association of Survivin and HBXIP comprising:
contacting Survivin and HBXIP in the presence or absence of a compound; and
detecting an altered association between Survivin and HBXIP, thereby determining whether said compound is an effective agent for altering association of Survivin with HBXIP.
2. The method of claim 1 , wherein Survivin and HBXIP associate in the presence of a compound and pro-Caspase-9.
3. The method of claim 1 , wherein the altered association between Survivin and HBXIP is detected by measuring the activation of pro-Caspase-9.
4. A method for identifying an effective agent that alters the association of Survivin and HBXIP comprising:
contacting Survivin and HBXIP under conditions that allow Survivin and HBXIP to associate in the presence of a compound, pro-Caspase-3, pro-Caspase-9, Apafl and cytochrome C; and
detecting an altered association between Survivin and HBXIP, thereby determining whether said compound is an effective agent for altering association of Survivin with HBXIP.
5. The method of claim 4 , wherein the altered association between Survivin and HBXIP is detected by measuring the activation of pro-Caspase-3.
6. The method of claim 5 , wherein the activation of pro-Caspase-3 is measured by monitoring the cleavage of caspase 3 substrate selected from the group consisting of DEVD-AFC, DEVD-pNA and DEVD-AMC.
7. A method for identifying an effective agent that alters the association of Survivin and HBXIP comprising:
contacting Survivin and HBXIP under conditions that allow Survivin and HBXIP to associate in the presence of a cell extract and a compound; and
detecting an altered association between Survivin and HBXIP, thereby determining whether said compound is an effective agent for altering association of Survivin with HBXIP.
8. The method of claim 7 , wherein said cell extract comprises pro-Caspase-9 and pro-Caspase-3.
9. The method of claim 7 , wherein the altered association between Survivin and HBXIP is detected by measuring the activation of pro-Caspase-3.
10. The method of claim 9 , wherein the activation of pro-Capase-3 is measured by monitoring the cleavage of a caspase 3 substrate selected from the group consisting of DEVD-AFC, DEVD-pNA and DEVD-AMC.