Targeting proteins for degradation
View Patent ↗Embodiments provided here include recombinant polypeptides, termed degradons, comprising a target binding domain and a proteasome-binding domain. Degradons of the embodiments are able to selectively target and degrade proteins bound by the target-binding domain, such as proteins associated with disease. Vectors encoding degradons and methods of treating disease with degradons and degradon expression vectors are likewise provided.
1. A recombinant polypeptide that binds to a target polypeptide, the recombinant polypeptide comprising a target-binding domain and a proteasome-binding domain that comprises a ubiquitin-like domain, wherein said target binding domain comprises an antibody or a fragment thereof that binds to the target polypeptide.
2. The polypeptide of claim 1 , wherein the target polypeptide is a polypeptide associated with a disease.
3. The polypeptide of claim 1 , wherein the target polypeptide is not a reporter protein; wherein the target-binding domain does not bind to the Huntingtin protein (HTT); or wherein the target-binding domain does not bind to protein having a poly-Q sequence.
4. The polypeptide of claim 1 , wherein the target-binding domain is position N-terminally relative to the proteasome binding domain.
5. The polypeptide of claim 1 , wherein the target-binding domain is position C-terminally relative to the proteasome binding domain.
6. The polypeptide of claim 1 , wherein the proteasome-binding domain comprises a domain from human Rad23b.
7. The polypeptide of claim 6 , wherein the proteasome-binding domain comprises amino acids 1-83 of human Rad23b.
8. The polypeptide of claim 1 , wherein the target-binding domain and the proteasome-binding domain are separated by a linker.
9. The polypeptide of claim 1 , further comprising a cell-penetrating peptide (CPP) sequence or a cellular receptor-binding sequence.
10. The polypeptide of claim 1 , wherein the antibody or antibody fragment comprises a monobody or scFv.
11. The polypeptide of claim 10 , wherein the antibody or antibody fragment comprises the HA4 monobody, Nsa1 monobody, and/or the Cs1 monobody.
12. The polypeptide of claim 1 , wherein the target-binding domain binds to a prion, a viral polypeptide, a disease-associated protein, a cellular polypeptide having a disease-associated mutation or the product of an oncogene.
13. The polypeptide of claim 12 , wherein the target-binding domain binds to the product of an oncogene.
14. The polypeptide of claim 12 , wherein the oncogene is Abl and/or Shp2.
15. The polypeptide of claim 12 , wherein the target binding domain binds to the SH2 domain of Abl, the N-terminal SH2 domain of Shp2, and/or C-terminal SH2 domain of Shp2.
16. The polypeptide of claim 12 , wherein the target-binding domain binds to misfolded beta-amyloid.
17. A nucleic acid molecule encoding a polypeptide according to claim 1 .
18. A pharmaceutical composition comprising: a polypeptide according to claim 1 .
19. The polypeptide of claim 1 , wherein the target binding domain comprises a monobody that binds to the target polypeptide.
20. The polypeptide of claim 1 , wherein the target binding domain comprises an scFv that binds to the target polypeptide.
21. The polypeptide of claim 1 , wherein the target binding domain comprises an antibody that binds to the target polypeptide.