Circular tandem repeat proteins
Circular handed alpha-helical repeat proteins are described. The repeat proteins have a number of uses as scaffolds for geometrically precise, arrayed presentation of cell-signaling or immune-related protein and peptide epitopes, as well as numerous other therapeutic, diagnostic, and nanotechnological uses.
1. A protein having the formula: (d-a-b-x-y) n, (a-d-b-x-y) n , (a-b-d-x-y) n , (a -b-x-d-y) n or (a-b-x-y-d) n herein b and y represent linkers, a represents an amino acid sequence that forms an alpha (α) helix, x represents an amino acid sequence that forms a second α helix, d represents a functional domain, and the protein is handed, wherein n is 2 or more.
2. The protein of claim 1 , wherein a and x are individually a sequence as set forth in SEQ ID NOs: 1-50, 124-129, 139, 140, 146, or 147 or a sequence having at least 98% sequence identity to SEQ ID NOs: 1-50, 124-129, 139, 140, 146, or 147.
3. The protein of claim 1 , wherein n is 3, 6, 9, 12, or 24.
4. The protein of claim 1 , wherein the N- and C-termini of the protein create a circular architecture.
5. The protein of claim 1 , wherein the functional domain comprises a cytokine, a Notch ligand, an immunogenic peptide, a peptide adjuvant, a single-chain class I MHC domain, or a small molecule ligand binding domain.
6. The protein of claim 1 , wherein the functional domain comprises SH2, SH3, IL-2, IL-3, IL-17c, single-chain MHC, the extracellular domain of the Delta-1 Notch protein ligand, Protein L, a protein having the sequence set forth in SEQ ID NO: 116, or a protein having the sequence set forth in SEQ ID NO: 115.
7. The protein of claim 1 , wherein the linkers comprise GBB linkers.
8. The protein of claim 7 , wherein the GBB linkers comprise GKS, GIT, GTT, GYS, GDK, GDE, NDK, GDR, GDL, or GIS.
9. The protein of claim 1 , wherein the protein is left-handed.
10. A circular, handed protein comprising α-helical structures wherein each α-helical structure comprises an outer α helix and an inner α helix joined by a flexible linker and wherein each α-helical structure has at least 90% sequence identity with an adjacent α-helical structure.
11. The circular, handed protein of claim 10 , further comprising a functional domain wherein the functional domain is inserted into the sequence of the protein between an outer a helix of the protein and an adjacent inner a helix of the protein.
12. The circular, handed protein of claim 11 , wherein the functional domain is a cytokine, a Notch ligand, an immunogenic peptide, a peptide adjuvant, a single- chain class I MHC domain, or a small molecule ligand binding domain.
13. The circular, handed protein of claim 11 , wherein the functional domain comprises SH2, SH3, IL-2, IL-3, IL-17c, single-chain MHC, the extracellular domain of the Delta-1 Notch protein ligand, Protein L, a protein having the sequence set forth in SEQ ID NO: 116, or a protein having the sequence set forth in SEQ ID NO: 115.
14. The circular, handed protein of claim 10 , wherein the linkers comprise a GBB linker.
15. The circular, handed protein of claim 14 , wherein the GBB linkers comprise GKS, GIT, GTT, GYS, GDK, GDE, NDK, GDR, GDL, or GIS.
16. An engineered, handed protein comprising alpha (α) helical structures wherein adjacent α helical structures are joined by a flexible linker, wherein each α-helical structure comprises an outer α helix and an inner α helix and wherein each α-helical structure has at least 95% sequence identity with an adjacent α-helical structure.
17. The engineered, handed protein of claim 16 , wherein the engineered, handed-protein is circular.
18. The engineered, handed protein of claim 16 , wherein the flexible linker comprises a GBB linker.
19. The engineered, handed protein of claim 18 , wherein the GBB linker comprises GKS, GIT, GTT, GYS, GDK, GDE, NDK, GDR, GDL, or GIS.
20. The engineered, handed protein of claim 19 , wherein the engineered protein comprises at least two GBB linkers.