Variants derived from ActRIIB
In certain aspects, the present invention provides compositions and methods for modulating (promoting or inhibiting) growth of a tissue, such as bone, cartilage, muscle, fat, and/or neuronal tissue. The present invention also provides methods of screening compounds that modulate activity of an ActRIIB protein and/or an ActRIIB ligand. The compositions and methods provided herein are useful in treating diseases associated with abnormal activity of an ActRIIB protein and/or an ActRIIB ligand.
1. A variant ActRIIB protein comprising an amino acid sequence that is at least 95% identical to the sequence of amino acids 29-109 of SEQ ID NO:2, wherein the protein comprises at least one N-X-S/T sequence at a position other than an endogenous N-X-S/T sequence of ActRIIB, wherein the at least one N-X-S/T sequence is at a position outside of the ligand-binding pocket, wherein the protein comprises a N at the position corresponding to position 24 of SEQ ID NO: 2 and a S or T at the position corresponding to position 26 of SEQ ID NO: 2 and wherein the variant ActRIIB protein inhibits signaling by myostatin and/or GDF11 in a cell-based assay.
2. The variant ActRIIB protein of claim 1 , wherein the protein comprises a R at the position corresponding to position 64 of SEQ ID NO:2.
3. The variant ActRIIB protein of claim 1 , wherein the protein comprises at least one additional alteration with respect to the sequence of SEQ ID NO:2, and wherein the at least one additional alteration is a conservative alteration positioned within the ligand binding pocket.
4. The variant ActRIIB protein of claim 1 , wherein the protein comprises at least one additional alteration with respect to the sequence of SEQ ID NO:2, and wherein the at least one additional alteration is an alteration at one or more positions selected from an amino acid corresponding to K74, R40, Q53, K55, F82 and L79 of SEQ ID NO:2.
5. The variant ActRIIB protein of claim 1 , wherein the protein comprises an amino acid sequence beginning at an amino acid corresponding to any of amino acids 22-25 of SEQ ID NO:2 and ending at an amino acid corresponding to any of amino acids 131, 133, or 134 of SEQ ID NO:2.
6. The variant ActRIIB protein of claim 1 , wherein the protein is a fusion protein further comprising a heterologous portion.
7. The variant ActRIIB protein of claim 6 , wherein the protein is a homodimer.
8. The variant ActRIIB protein of claim 6 , wherein the heterologous portion comprises a constant region from an IgG heavy chain.
9. The variant ActRIIB protein of claim 1 , wherein the protein comprises an amino acid sequence that is at least 98% identical to the sequence of amino acids 29-109 of SEQ ID NO:2.
10. The variant ActRIIB protein of claim 1 , wherein the protein comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:5.
11. The variant ActRIIB protein of claim 10 , wherein the protein has an N at the position corresponding to position 5 of SEQ ID NO:5.
12. The variant ActRIIB protein of claim 10 , wherein the protein has a D the position corresponding to position 60 of SEQ ID NO:5.
13. The variant ActRIIB protein of claim 10 , wherein the protein has an E at the position corresponding to position 60 of SEQ ID NO:5.
14. The variant ActRIIB protein of claim 10 , wherein the protein is a dimer.
15. The variant ActRIIB protein of claim 11 , wherein the protein is a dimer.
16. The variant ActRIIB protein of claim 12 , wherein the protein is a dimer.
17. The variant ActRIIB protein of claim 13 , wherein the protein is a dimer.
18. A pharmaceutical preparation comprising the protein of claim 1 .
19. The variant ActRIIB protein of claim 1 , wherein the protein comprises a K at the position corresponding to position 64 of SEQ ID NO:2.
20. The variant ActRIIB protein of claim 1 , wherein the protein comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:5.
21. The variant ActRIIB protein of claim 1 , wherein the protein comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO:5.
22. The variant ActRIIB protein of claim 1 , wherein the protein comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:12.
23. The variant ActRIIB protein of claim 1 , wherein the protein comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO:12.
24. The variant ActRIIB protein of claim 1 , wherein the protein comprises one or more modified amino acid residues selected from: a glycosylated amino acid, a PEGylated amino acid, a farnesylated amino acid, an acetylated amino acid, a biotinylated amino acid, and an amino acid conjugated to a lipid moiety.
25. The variant ActRIIB protein of claim 6 , wherein the heterologous portion comprises an Fc domain.
26. The variant ActRIIB protein of claim 25 , wherein the Fc domain comprises the amino acid sequence of SEQ ID NO:13.
27. The variant ActRIIB protein of claim 6 , wherein the fusion protein further comprises a linker domain positioned between the ActRIIB polypeptide and the heterologous polypeptide.
28. The variant ActRIIB protein of claim 1 , wherein the protein inhibits signaling by myostatin in a cell-based assay.
29. The variant ActRIIB protein of claim 1 , wherein the protein inhibits signaling by GDF11 in a cell-based assay.