Sox-based kinase sensor
Peptidyl sensors comprise a metal-binding peptide and one or two kinase recognition sequences with a hydroxyamino acid that can be phosphorylated in the presence of a kinase.
1. A peptide comprising an amino acid residue, a side chain of the amino acid residue reacted with a compound of structure (I):
wherein R 1 is hydroxy, amino, or thiol;
R 2 and R 3 are each independently hydrogen, an electron-withdrawing group, or —SO 2 X;
each R 4 is independently hydrogen, an electron-withdrawing group or —SO 2 X, wherein R 4 can substitute any open valence of any ring within structure (I);
X is —OR′ or —NR′R″;
Y is a moiety that reacts with the side chain of an amino acid residue of the peptide to produce a thioether linkage between structure (I) and the side chain of the amino acid residue of the peptide;
R′ and R″ are each independently hydrogen or C 1-6 alkyl;
n is 0, 1, 2 or 3; and
m is 1, 2 or 3, 4, and
wherein at least one of R 2 , R 3 or R 4 is —SO 2 X.
2. The peptide of claim 1 , wherein the amino acid residue is a cysteine.
3. The peptide of claim 1 , wherein the peptide further comprises at least one kinase recognition sequence.
4. The peptide of claim 1 , wherein n is 0.
5. The peptide of claim 1 , wherein R 1 is hydroxy.
6. The peptide of claim 1 , wherein R 2 and R 3 are each hydrogen.
7. The peptide of claim 1 , wherein one R 4 is para to R 1 .
8. The peptide of claim 7 , wherein R 4 is —SO 2 X.
9. A peptide comprising an amino acid residue, wherein a side chain of the amino acid residue is connected to structure (I):
wherein R 1 is hydroxy, amino, or thiol;
R 2 and R 3 are each independently hydrogen, an electron-withdrawing group, or —SO 2 X;
each R 4 is independently hydrogen, an electron-withdrawing group or —SO 2 X, wherein R 4 can substitute any open valence of any ring within structure (I);
X is —OR′ or —NR′R″;
Y is a thioether linkage, a disulfide linkage, or an amide linkage connecting structure (I) to the side chain of the amino acid residue;
R′ and R″ are each independently hydrogen or C 1-6 alkyl;
n is 0, 1, 2 or 3; and
m is 1, 2 or 3, 4, and
wherein at least one of R 2 , R 3 or R 4 is —SO 2 X.
10. The peptide of claim 9 , wherein the amino acid residue is a cysteine.
11. The peptide of claim 9 , wherein the peptide further comprises at least one kinase recognition sequence.
12. The peptide of claim 9 , wherein n is 0.
13. The peptide of claim 9 , wherein R 1 is hydroxy.
14. The peptide of claim 9 , wherein R 2 and R 3 are each hydrogen.
15. The peptide of claim 9 , wherein one R 4 is para to R 1 .
16. The peptide of claim 15 , wherein R 4 is —SO 2 X.
17. The peptide of claim 15 , wherein Y is a thioether linkage.
18. The peptide of claim 15 , wherein Y is a disulfide linkage.
19. The peptide of claim 15 , wherein Y is an amide linkage.