Self-assembling peptide incorporating modifications and methods of use thereof
The invention provides a self-assembling peptide comprising (a) a first amino acid domain that mediates self-assembly, wherein the domain comprises alternating hydrophobic and hydrophilic amino acids that are complementary and structurally compatible and self-assemble into a macroscopic structure when present in unmodified form; and (b) a second amino acid domain that does not self-assemble in isolated form. In certain embodiments of the invention the second amino acid domain comprises a biologically active peptide motif, e.g., a peptide motif found in a naturally occurring protein, or a target site for an interaction with a biomolecule. In certain embodiments of the invention the naturally occurring protein is a component of the extracellular matrix, e.g., a component of the basement membrane. The invention further provides scaffolds comprising the self-assembling peptides and methods of using the scaffolds including for cell culture, tissue engineering, and tissue repair.
1. A self-assembling peptide comprising:
(a) a first amino acid domain that mediates self-assembly, wherein the domain comprises a sequence of alternating hydrophobic and hydrophilic amino acids that are complementary and structurally compatible, and that self-assemble into a macroscopic structure when present in unmodified form, which sequence is selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 18; and
b) a second amino acid domain that does not self-assemble in isolated form,
wherein the first and second amino acid domains are joined by a single peptide bond;
wherein the peptide self-assembles when present as the only peptide in a solution; and
wherein the total number of amino acids comprising the self-assembling peptide is less than 36 amino acids.
2. The self-assembling peptide of claim 1 wherein the second amino acid domain comprises a biologically active peptide motif or a target site for an interaction with a biomolecule.
3. The self-assembling peptide of claim 2 , wherein the sequence of the second amino acid domain comprises a sequence selected from the group consisting of: SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, and SEQ ID NO: 37.
4. The self-assembling peptide of claim 1 , wherein the peptide is linear.
5. The self-assembling peptide of claim 1 , wherein the first amino acid domain comprises multiple RADA (SEQ ID NO: 4) peptide subunits.
6. The self-assembling peptide of claim 1 , wherein the total number of amino acids comprising the second amino acid domain is less than 20 amino acids.
7. The self-assembling peptide of claim 1 , wherein the total number of amino acids comprising the second amino acid domain is less than 12 amino acids.
8. The self-assembling peptide of claim 1 , wherein the total number of amino acids comprising the second amino acid domain is less than 7 amino acids.
9. The self-assembling peptide of claim 1 , wherein the total number of amino acids comprising the second amino acid domain is less than 5 amino acids.
10. The self-assembling peptide of claim 1 , wherein the first amino acid domain comprises RADA16-I peptide.
11. The self-assembling peptide of claim 1 , wherein the peptide self-assembles to form a hydrogel.
12. The self-assembling peptide of claim 1 , further comprising a third amino acid domain joined by a peptide bond to the first amino acid domain.
13. The self-assembling peptide of claim 1 , further comprising a third amino acid domain joined by a peptide bond to the second amino acid domain.
14. A self-assembling peptide comprising:
(a) a first amino acid domain that mediates self-assembly, wherein the domain comprises a sequence of alternating hydrophobic and hydrophilic amino acids that are complementary and structurally compatible, and that self-assemble into a macroscopic structure when present in unmodified form, which sequence is selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 18; and
b) a second amino acid domain that does not self-assemble in isolated form,
wherein the first and second amino acid domains are joined by a linker domain,
wherein the peptide self-assembles when present as the only peptide in a solution; and
wherein the total number of amino acids comprising the self-assembling peptide is less than about 36 amino acids.
15. The self-assembling peptide of claim 14 , wherein the linker domain is between 1 and 4 amino acids.
16. The self-assembling peptide of claim 14 , wherein the linker domain is between 1 and 4 glycine residues.
17. The self-assembling peptide of claim 14 , wherein the linker domain comprises one or more glycine residues.
18. The self-assembling peptide of claim 14 , wherein the first amino acid domain comprises multiple RADA (SEQ ID NO: 4) peptide subunits.
19. The self-assembling peptide of claim 14 , wherein the total number of amino acids comprising the second amino acid domain is less than 20 amino acids.
20. The self-assembling peptide of claim 14 , wherein the total number of amino acids comprising the second amino acid domain is less than 12 amino acids.
21. The self-assembling peptide of claim 14 , wherein the total number of amino acids comprising the second amino acid domain is less than 7 amino acids.
22. The self-assembling peptide of claim 14 , wherein the total number of amino acids comprising the second amino acid domain is less than 5 amino acids.
23. The self-assembling peptide of claim 14 , wherein the first amino acid domain comprises RADA16-I peptide.
24. The self-assembling peptide of claim 14 , wherein the peptide self-assembles to form a hydrogel.
25. The self-assembling peptide of claim 14 , further comprising a third amino acid domain joined by a peptide bond to the first amino acid domain.
26. The self-assembling peptide of claim 14 , further comprising a third amino acid domain joined by a peptide bond to the second amino acid domain.
27. The self-assembling peptide of claim 2 , wherein the sequence of the second amino acid domain comprises a sequence selected from the group consisting of: SEQ ID NO: 31, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, and SEQ ID NO: 37.
28. The self-assembling peptide of claim 14 , wherein the sequence of the second amino acid domain comprises a sequence selected from the group consisting of: SEQ ID NO: 31, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, and SEQ ID NO: 37.
29. The self-assembling peptide of claim 1 having the sequence AcN-RYVVLPR-GG-(RADA) 4 -CONH 2 (SEQ ID NO: 43).
30. A self-assembling peptide comprising:
(a) a first amino acid domain that mediates self-assembly, wherein the domain comprises alternating hydrophobic and hydrophilic amino acids that are complementary and structurally compatible, and that self-assemble into a macroscopic structure when present in unmodified form, which sequence is SEQ ID NO: 4 (RAD 16-I); and
(b) a second amino acid domain, wherein the sequence of the second amino acid domain comprises SEQ ID NO: 35 (RYVVLPR); and
Wherein the peptide forms a beta sheet upon self-assembly.