Protein-based adhesives
Protein-based adhesives. In one embodiment of the present disclosure, an elastin-like polypeptide has a sequence LDGTL-(PGX 1 GVPGKGVPGX 2 GVPGX 1 GVPGX 3 GVPGX 2 GV) n -PVADRGMRLE, wherein each X 1 is selected from the group consisting of tyrosine (Y), dihydroxyphenylalanine (DOPA), and 3,4,5-trihydroxyphenylalanine (TOPA), wherein each X 2 is selected from the group consisting of valine (V), Y, DOPA, and TOPA, wherein each X 3 is selected from the group consisting of glutamic acid (E) and lysine (K), and wherein n is at or between 6 and 10 or higher or lower.
1. An elastin-like polypeptide comprising LDGTL-(PGX 1 GVPGKGVPGX 2 GVPGX 1 GVPGX 3 GVPGX 2 GV) n -PVADRGMRLE (SEQ ID NO: 2), wherein:
each X 1 is independently selected from the group consisting of tyrosine (Y), dihydroxyphenylalanine (DOPA), and 3,4,5-trihydroxyphenylalanine (TOPA);
each X 2 is independently selected from the group consisting of valine (V), Y, DOPA, and TOPA,
X 3 is independently selected from the group consisting of glutamic acid (E) and lysine (K), and
n is an integer from 6 to 10.
2. The elastin-like polypeptide of claim 1 , wherein n is 8.
3. The elastin-like polypeptide of claim 1 , comprising DDDDK-LDGTL-(PGX 1 GVPGKGVPGX 2 GVPGX 1 GVPGX 3 GVPGX 2 GV) n -PVADRGMRLE (SEQ ID NO: 7), wherein:
each X 1 is independently selected from the group consisting of Y, DOPA, and TOPA;
each X 2 is independently selected from the group consisting of V, Y, DOPA, and TOPA;
X 3 is independently selected from the group consisting of E and K;
n is an integer from 6 to 10; and
the sequence N-terminal to LDTGTL (SEQ ID NO: 10) in SEQ ID NO: 7 comprises a cleavage site.
4. The elastin-like polypeptide of claim 3 , comprising HHHHHHH-DDDDK-LDGTL-(PGX 1 GVPGKGVPGX 2 GVPGX 1 GVPGX 3 GVPGX 2 GV) n -PVADRGMRLE (SEQ ID NO: 4), wherein:
each X 1 is independently selected from the group consisting of Y, DOPA, and TOPA;
each X 2 is independently selected from the group consisting of V, Y, DOPA, and TOPA;
X 3 is independently selected from the group consisting of E and K;
n is an integer from 6 to 10; and
the repeating histidine residues at the N-terminus of SEQ ID NO: 4 are a His-tag.
5. The elastin-like polypeptide of claim 4 , comprising M-MASMTGGQQMG-HHHHHHH-DDDDK-LDGTL-(PGX 1 GVPGKGVPGX 2 GVPGX 1 GVPGX 3 GVPGX 2 GV) n -PVADRGMRLE (SEQ ID NO: 6), wherein:
each X 1 is independently selected from the group consisting of Y, DOPA, and TOPA;
each X 2 is independently selected from the group consisting of V, Y, DOPA, and TOPA;
X 3 is independently selected from the group consisting of E and K;
n is an integer from 6 to 10; and
the sequence N-terminal to the His-tag in SEQ ID NO: 6 is a T7 tag.
6. The elastin-like polypeptide of claim 1 , having a lower critical solution temperature (LCST) at or between 25° C. and 37° C.
7. The elastin-like polypeptide of claim 1 , wherein X 1 is Y, X 2 is V, and X 3 is K.
8. The elastin-like polypeptide of claim 1 , wherein at least one X 1 is DOPA or TOPA.
9. The elastin-like polypeptide of claim 1 , wherein n is 8 and X 1 is Y.
10. The elastin-like polypeptide of claim 1 , capable of adhering to a substrate when applied to said substrate under wet conditions.
11. The elastin-like polypeptide of claim 1 , wherein X 2 is V and X 3 is K.
12. An elastin-like polypeptide comprising MSKGPGVDGTL-(PGX 1 GVPGKGVPGX 2 GVPGX 1 GVPGX 3 GVPGX 2 GV) n -PVADRGMRLE (SEQ ID NO: 3), wherein:
each X 1 is independently selected from the group consisting of Y, DOPA, and TOPA;
each X 2 is independently selected from the group consisting of V, Y, DOPA, and TOPA;
X 3 is independently selected from the group consisting of E and K; and
n is an integer from 6 to 10.
13. An elastin-like polypeptide comprising VPGXG repeated at least six times within the elastin-like polypeptide (SEQ ID NO: 8), wherein each X is independently selected from the group consisting of E, K, V, Y, DOPA, and TOPA and at least one X is DOPA or TOPA.
14. A method of generating a polypeptide configured for wet adhesion, comprising the steps of:
providing an initial polypeptide comprising SEQ ID NO: 9; and
dissolving the initial polypeptide in a buffer comprising a tyrosinase to form a mixture so that least one tyrosine of the initial polypeptide is converted to dihydroxyphenylalanine (DOPA) within the mixture.
15. The method of claim 14 , wherein the step of dissolving further comprises converting at least one DOPA to 3,4,5-trihydroxyphenylalanine (TOPA).
16. The method of claim 14 , further comprising the step of:
adding an acid to the mixture after a first period of time has elapsed so to cease further conversion of tyrosine to DOPA.