Inhibitor of apoptosis proteins and nucleic acids and methods for making and using them
View Patent ↗The invention provides polypeptides comprising inhibitor of apoptosis protein (IAP) family members, such as BmIAP initially derived from Bombyx mori BmN cells, and nucleic acids encoding them, and methods for making and using these compositions, including their use for inhibiting apoptosis.
1. A method for identifying an agent that can modulate the activity of a polypeptide, wherein the polypeptide comprises a sequence having at least 95% sequence identity to SEQ ID NO:2 and is capable inhibiting apoptosis in a cell, comprising: (a) contacting a cell expressing the polypeptide recombinantly in the presence and absence of a test agent before, during or after inducing apoptosis in the cell; and (b) measuring the amount or degree of polypeptide activity in the cell in the presence and absence of the test agent, wherein an increase or decrease in the amount or degree of polypeptide activity in the cell in the presence of the test agent identifies the test agent as a modulator of the polypeptide's activity.
2. The method of claim 1 , wherein the cell is an insect cell.
3. The method of claim 2 , wherein the cell is a Bombyx mori cell.
4. The method of claim 1 , wherein the cell is a plant cell.
5. The method of claim 1 , wherein the cell is a mammalian cell.
6. The method of claim 1 , wherein the cell is a yeast cell.
7. The method of claim 1 , wherein the degree of polypeptide activity in the cell is determined by measuring the amount or degree of apoptosis in the cell.
8. The method of claim 1 , wherein the degree of polypeptide activity in the cell is determined by measuring the amount or degree of caspase protease activity in the cell.
9. The method of claim 1 , wherein the degree of polypeptide activity in the cell is determined by measuring the amount or degree of DNA fragmentation in the cell.
10. The method of claim 1 , wherein the degree of polypeptide activity in the cell is determined by measuring the amount or degree of cleavage of substrates of caspases in the cell.