Designer ligands of TGF-β superfamily
The present disclosure relates to chimeric polypeptide having TGF-beta activity, nucleic acids encoding the polypeptides, and host cells for producing the polypeptides.
1. A recombinant TGF-β ligand polypeptide having at least 95% sequence identity to an amino acid sequence comprising
a first segment of amino acid residues comprising amino acid residues 1 to 32 of SEQ ID NO:2 (“1b”);
a second segment of amino acid residues comprising amino acid residues 30 to 45 of SEQ ID NO:5 (“2a”);
a third segment of amino acid residues comprising amino acid residues 46 to x 1 of SEQ ID NO:5 (“3a”), or amino acid residues x 2 to x 3 of SEQ ID NO:2 (“3b”),
wherein:
x 1 is residue 61, 62, 63, or 64 of SEQ ID NO:5,
x 2 is residue 45, 46, 47, or 48 of SEQ ID NO:2, and
x 3 is residue 65, 66, 67, or 68 of SEQ ID NO:2; and
a fourth segment of amino acid residues comprising amino acid residues x 4 to x 5 of SEQ ID NO:5 (“4a”) or amino acid residues x 6 to x 7 of SEQ ID NO:2 (“4b”),
wherein:
x 4 is x 1 of SEQ ID NO: 5 if the third segment comprises 3b, or x 4 is x 1 +1 of SEQ ID NO: 5 if the third segment comprises 3a,
x 5 is residue 78, 79, 80, 81, 82, 83 or 84 of SEQ ID NO:5,
x 6 is x 3 of SEQ ID NO: 2 if the third segment comprises 3a, or x 6 is x 3 +1 of SEQ ID NO: 2 if the third segment comprises 3b,
x 7 is residue 76, 77, 78, 79, 80, 81 or 82 of SEQ ID NO:2, and;
a fifth segment of amino acid residues comprising amino acid residues x 8 to x 9 of SEQ ID NO:5 (“5a”) or amino acid residues x 10 to x 11 of SEQ ID NO:2 (“5b”),
wherein:
x 8 is x 5 of SEQ ID NO: 5 if the fourth segment comprises 4b, or x 8 is x 5 +1 of SEQ ID NO: 5 if the fourth segment comprises 4a,
x 9 is residue 90, 91, 92, 93, 94, 95 or 96 of SEQ ID NO:5,
x 10 is x 7 of SEQ ID NO: 2 if the fourth segment comprises 4a, or x 10 is x 7 +1 of SEQ ID NO: 2 if the fourth segment comprises 4b,
x 11 is residue 88, 89, 90, 91, 92, 93, or 94 of SEQ ID NO:2, and
a sixth segment comprising amino acid residues x 12 to x 13 of SEQ ID NO:5 (“6a”) or amino acid residues x 14 to x 15 of SEQ ID NO:2 (“6b”),
wherein:
x 12 is x 9 of SEQ ID NO: 5 if the fifth segment comprises 5b, or x 12 is x 9 +1 of SEQ ID NO: 5 if the fifth segment comprises 5a,
x 13 is residue 114, 115, or 116 or SEQ ID NO:5,
x 14 is x 11 of SEQ ID NO: 2 if the fifth segment comprises 5a, or x 14 is x 11 +1 of SEQ ID NO: 2 if the fifth segment comprises 5b,
x 15 is residue 112, 113, or 114 or SEQ ID NO:2;
wherein the recombinant TGF-β ligand polypeptide activates cell proliferation or activity associated with SMAD pathway.
2. The recombinant polypeptide of claim 1 , wherein
the third segment comprises amino acid residues x 2 to x 3 of SEQ ID NO:2 (“3b”);
the fourth segment comprises amino acid residues x 6 to x 7 of SEQ ID NO:2 (“4b”), wherein x 6 is x 3 +1 of SEQ ID NO: 2;
the fifth segment comprises amino acid residues x 8 to x 9 of SEQ ID NO:5 (“5a”), wherein x 8 is x 5 of SEQ ID NO: 5; and
the sixth segment comprises amino acid residues x 12 to x 13 of SEQ ID NO:5 (“6a”), wherein x 12 is x 9 +1 of SEQ ID NO: 5.
3. The recombinant polypeptide of claim 1 , wherein
the third segment comprises amino acid residue 46 to x 1 of SEQ ID NO:5 (“3a”);
the fourth segment comprises amino acid residues x 4 to x 5 of SEQ ID NO:5 (“4a”), wherein x 4 is x 1 +1 of SEQ ID NO: 5;
the fifth segment comprises amino acid residues x 8 to x 9 of SEQ ID NO:5 (“5a”), wherein x 8 is x 5 +1 of SEQ ID NO: 5; and
the sixth segment comprises amino acid residues x 12 to x 13 of SEQ ID NO:5 (“6a”), wherein x 12 is x 9 +1 of SEQ ID NO: 5.
4. The recombinant polypeptide of claim 1 , wherein
the third segment comprises amino acid residues x 2 to x 3 of SEQ ID NO:2 (“3b”);
the fourth segment comprises amino acid residues x 6 to x 7 of SEQ ID NO:2 (“4b”), wherein x 6 is x 3 +1 of SEQ ID NO: 2;
the fifth segment comprises amino acid residues x 10 to x 11 of SEQ ID NO:2 (“5b”), wherein x 10 is x 7 +1 of SEQ ID NO: 2; and
the sixth segment comprises amino acid residues x 12 to x 13 of SEQ ID NO:5 (“6a”), wherein x 12 is x 9 of SEQ ID NO: 5.
5. The polypeptide of claim 1 , wherein the polypeptide comprises a sequence selected from the group consisting of SEQ ID NO: 17, 19, 23, 25, 27, 29, 31, and 33.
6. The recombinant polypeptide of claim 1 , wherein said polypeptide comprises the sequence as set forth in SEQ ID NO: 31.
7. The recombinant polypeptide of claim 1 , wherein said polypeptide comprises the sequence as set forth in SEQ ID NO: 17.
8. The recombinant polypeptide of claim 1 , wherein said polypeptide comprises the sequence as set forth in SEQ ID NO: 29.
9. The recombinant polypeptide of claim 1 , wherein said polypeptide has at least 95% sequence identity to the sequence as set forth in SEQ ID NO: 31.
10. The recombinant TGF-β ligand polypeptide of claim 1 , wherein the recombinant TGF-β ligand polypeptide has at least 97% sequence identity to the amino acid sequence comprising the first, the second, the third, the fourth, the fifth, and the sixth segments.
11. The recombinant TGF-β ligand polypeptide of claim 1 , wherein the recombinant TGF-β ligand polypeptide has at least 98% sequence identity to the amino acid sequence comprising the first, the second, the third, the fourth, the fifth, and the sixth segments.
12. The recombinant TGF-β ligand polypeptide of claim 1 , wherein the recombinant TGF-β ligand polypeptide has 100% sequence identity to the amino acid sequence comprising the first, the second, the third, the fourth, the fifth, and the sixth segments.
13. A polynucleotide comprising a sequence selected from the group consisting of SEQ ID NOs: 16, 18, 22, 24, 26, 28, 30, and 32.
14. The polynucleotide of claim 13 , wherein said polynucleotide comprises the sequence as set forth in SEQ ID NO: 30.
15. The polynucleotide of claim 13 , wherein said polynucleotide comprises the sequence as set forth in SEQ ID NO: 16.
16. The polynucleotide of claim 13 , wherein said polynucleotide comprises the sequence as set forth in SEQ ID NO: 28.