Polynucleotides encoding soluble, stable forms of human double minute 2 polypeptides
View Patent ↗The present invention discloses modified Hdm2 proteins that are soluble. In addition, the present invention discloses nucleic acids that encode the modified Hdm2 proteins of the present invention. The invention also provides crystals of modified Hdm2 proteins that are suitable for X-ray crystallization analysis. The present invention also discloses methods of using the modified Hdm2 proteins and crystals thereof to identify, select and/or design compounds that may be used as anticancer agents. The present invention further discloses compounds that bind to modified Hdm2 proteins in protein-ligand complexes.
1. A nucleic acid encoding a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 10 and SEQ ID NO: 12.
2. The nucleic acid according to claim 1 comprising a nucleotide sequence selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 9, and SEQ ID NO: 11.
3. An expression vector comprising the nucleic acid according to claim 2 , wherein said expression vector further comprises a transcriptional control sequence operatively linked to the nucleic acid.
4. A host cell comprising the expression vector according to claim 3 .
5. A method for producing a modified Hdm2 protein comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 10 and SEQ ID NO: 12, comprising culturing the host cell according to claim 4 in a culture medium under conditions in which the nucleic acid encoding the modified Hdm2 protein is expressed, thereby producing the modified Hdm2 protein.
6. The method according to claim 5 wherein the host cell is an Escherichia coli cell.
7. A method for obtaining a purified modified Hdm2 protein comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 6, SEQ ID NO: 10 and SEQ ID NO: 12, comprising culturing the host cell of claim 4 in a culture medium under conditions in which the nucleic acid encoding the modified Hdm2 protein is expressed to thereby produce the modified Hdm2 protein; and purifying the modified Hdm2 protein from the culture medium.