Kinase substrates
View Patent ↗Tyrosine kinase substrates are described herein that are phosphorylated by many and diverse tyrosine kinases, and are chemically stable relative to co-polymers of poly-EY or poly-EAY having random molecular weights in the range of 20-50 kDa. Tyrosine kinase substrate peptides are provided according to embodiments described herein which include an isolated tyrosine kinase substrate peptide having molecular weight in the range of about 0.5 kD-10 kD. Tyrosine kinase substrate peptides are provided according to embodiments described herein having no more than 50 amino acids. The peptides include 2-25 phosphorylation modules and each phosphorylation module has 2-3 amino acid residues.
1. An isolated tyrosine kinase substrate peptide, comprising:
an amino acid sequence selected from the group consisting of: AYE AYE AYE K EYA EYA EYA K AYA EYE (SEQ ID NO. 1); AYE AYE AYE AYA EYE (SEQ ID NO.2); C AYE AYE AYE K EYA EYA EYA K AYA EYE R (SEQ ID NO. 3); CGG E AYE AYE AYE AYA EYE ARR (SEQ ID NO.4); and AYE AYE AYE B EYA EYA EYA B AYA EYE (SEQ ID NO. 5), where each B is a basic amino acid residue independently selected from histidine, lysine and arginine.
2. The tyrosine kinase substrate peptide of claim 1 , wherein the peptide comprises at least one basic amino acid residue.
3. The tyrosine kinase substrate peptide of claim 2 , the at least one basic amino acid residue is selected from the group consisting of: histidine, lysine and arginine.
4. The tyrosine kinase substrate peptide of claim 1 , wherein the peptide has a net positive charge.
5. The tyrosine kinase substrate peptide of claim 1 , wherein the peptide comprises a detectable label.
6. The tyrosine kinase substrate peptide of claim 1 , wherein the detectable label is a FRET acceptor.
7. A plurality of tyrosine kinase substrate peptides, each peptide of the plurality comprising:
an amino acid sequence selected from the group consisting of: AYE AYE AYE K EYA EYA EYA K AYA EYE (SEQ ID NO. 1); AYE AYE AYE AYA EYE (SEQ ID NO.2); C AYE AYE AYE K EYA EYA EYA K AYA EYE R (SEQ ID NO. 3); CGG E AYE AYE AYE AYA EYE ARR (SEQ ID NO.4) and AYE AYE AYE B EYA EYA EYA B AYA EYE (SEQ ID NO. 5), where each B is a basic amino acid residue independently selected from histidine, lysine and arginine.
8. The plurality of tyrosine kinase substrate peptides of claim 7 , wherein each peptide has a net positive charge.
9. A kit for detection of tyrosine kinase activity, comprising:
a tyrosine kinase substrate peptide wherein the tyrosine kinase substrate peptide comprises an amino acid sequence selected from the group consisting of: AYE AYE AYE K EYA EYA EYA K AYA EYE (SEQ ID NO. 1); AYE AYE AYE AYA EYE (SEQ ID NO.2); C AYE AYE AYE K EYA EYA EYA K AYA EYE R (SEQ ID NO. 3); CGG E AYE AYE AYE AYA EYE ARR (SEQ ID NO.4) and AYE AYE AYE B EYA EYA EYA B AYA EYE (SEQ ID NO. 5), where each B is a basic amino acid residue independently selected from histidine, lysine and arginine.
10. The kit for detection of tyrosine kinase activity of claim 9 , wherein the peptide comprises a detectable label.
11. The kit for detection of tyrosine kinase activity of claim 10 , wherein the detectable label is a FRET acceptor.
12. The kit for detection of tyrosine kinase activity of claim 10 , wherein the N-terminus of the peptide comprises the detectable label.
13. The kit for detection of tyrosine kinase activity of claim 9 , wherein the peptide comprises at least one basic amino acid residue.
14. The kit for detection of tyrosine kinase activity of claim 13 , the at least one basic amino acid residue is selected from the group consisting of: histidine, lysine and arginine.
15. The kit for detection of tyrosine kinase activity of claim 9 , wherein the peptide has a net positive charge.