HDM2 Polypeptides
The present invention relates to HDM2 polypeptides and mutants thereof which are complexed with various compounds, e.g., HDM2 inhibitors.
1. A complex comprising an isolated polypeptide comprising:
(i) amino acids 17-125 of human HDM2 as set forth in SEQ ID NO: 1;
(ii) amino acids 17-125 of human HDM2 as set forth in SEQ ID NO: 1 comprising an F55Y mutation;
(iii) amino acids 17-125 of human HDM2 as set forth in SEQ ID NO: 1 comprising a Y76H mutation; or
(iv) amino acids 17-125 of human HDM2 as set forth in SEQ ID NO: 1 comprising an F55Y/Y76H double mutation; complexed with a compound.
2. The complex of claim 1 wherein said HDM2 amino acids 17-125 are:
SQIPASEQETLVRPKPLLLKLLKSVGAQKDTYTMKEVLYYLGQYIMTKRLYDEKQQHIV HCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSEN (SEQ ID NO: 2).
3. A method for making the complex of claim 1 comprising contacting said polypeptide and said compound.
4. A composition comprising ammonium sulfate and a complex of claim 1 comprising:
(a) amino acids 17-125 of human HDM2 as set forth in SEQ ID NO: 1;
(b) amino acids 17-125 of human HDM2 as set forth in SEQ ID NO: 1 comprising an F55Y mutation;
(c) amino acids 17-125 of human HDM2 as set forth in SEQ ID NO: 1 comprising a Y76H mutation; or
(d) amino acids 17-125 of human HDM2 as set forth in SEQ ID NO: 1 comprising an F55Y/Y76H double mutation; complexed with a compound.
5. The composition of claim 4 wherein said HDM2 amino acids 17-125 are:
SQIPASEQETLVRPKPLLLKLLKSVGAQKDTYTMKEVLYYLGQYIMTKRLYDEKQQHIV HCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSEN (SEQ ID NO: 2).
6. The composition of claim 4 wherein the polypeptide is present at a concentration of at least 10 mg/ml.
7. The composition of claim 4 having a temperature of about 22° C. or about 4° C.
8. The composition of claim 4 wherein the ammonium sulfate is at a concentration of about 0.9 M.