Interaction between the VHL tumor suppressor and hypoxia inducible factor, and assay methods relating thereto
View Patent ↗The invention relates to the finding that the VHL tumour suppressor protein regulates hypoxia inducible factor α subunits, by targeting HIF α for destruction in normoxic, but not hypoxic cells. The invention provides assays for modulators of this interaction, and peptides based upon HIF α subunit sequences which may modulate this interaction.
1. An isolated polypeptide which consists of from 8 to 7 amino acids, said polypeptide being capable of forming a complex with VHL, whose sequence is found in region 549-652 of human HIF-1 α (SEQ ID NO: 18).
2. The isolated polypeptide of claim 1 wherein said sequence is found in region 549-572 of human HIF-1 α (SEQ ID NO: 18).
3. The polypeptide of claim 1 fused to a membrane translocation sequence.
4. The polypeptide of claim 1 for use in a method of inhibiting the interaction of an HIF α subunit with VHL.
5. An isolated polypeptide which consists of from 8 to 27 amino acids, said polypeptide being capable of forming a complex with VHL, and characterized by the presence of a sequence selected from the group consisting of:
LAPYIPMD; SEQ ID NO: 1;
LAPYISMD; SEQ ID NO: 2;
LLPYIPMD; SEQ ID NO: 3;
LVPYIPMD; SEQ ID NO: 4;
IAPYIPMD; SEQ ID NO: 5;
IAPYIPME; SEQ ID NO: 6; and
LVPYISMD; SEQ ID NO: 7.
6. An isolated polypeptide which consists of from 19 to 27 amino acids, said polypeptide being capable of forming a complex with VHL, wherein said polypeptide comprises the sequence DLDLEMLAPYIPMDDDFQL (SEQ ID NO: 8); and variants thereof in which there are from 1 to 4 substitutions.
7. The polypeptide of claim 5 fused to a membrane translocation sequence.
8. An isolated polypeptide comprising multiple copies of peptides, wherein said copies of peptides are independently chosen from:
peptides which consist of from 8 to 27 amino acids, said peptides being capable of forming a complex with VHL, whose sequence is found in region 549-652 of human HIF-1 α (SEQ ID NO: 18), and
peptides which consist of from 8 to 27 amino acids, said peptides being capable of forming a complex with VHL, and characterized by the presence of a sequence selected from the group consisting of:
LAPYIPMD; SEQ ID NO: 1;
LAPYISMD; SEQ ID NO: 2;
LLPYIPMD; SEQ ID NO: 3;
LVPYIPMD; SEQ ID NO: 4;
IAPYIPMD; SEQ ID NO: 5;
IAPYIPME; SEQ ID NO: 6; and
LVPYISMD; SEQ ID NO: 7.