Degradation resistant peptide based biosensors
AssigneePurdue Research Foundation
InventorsLaurie L Parker, Robert John Schuster
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Abstract
Disclosed are compositions and methods for measuring tyrosine kinase activity using degradation resistant biosensors comprising a reporter sequence, a target sequence, and a transduction sequence. In certain embodiments, the invention includes a biosensor comprising a peptide comprising a reporter sequence, i.e., an amino acid sequence including a tyrosine residue that can be phosphorylated by a tyrosine kinase. In certain embodiments, a biosensor comprises a reporter sequence configured to resist degradation from proteases, a targeting sequence coupled to the reporter sequence and configured to bind to a protein interaction domain of a kinase, and a transduction sequence coupled to the targeting sequence.
Claims15
A biosensor comprising:
- a reporter sequence configured to resist degradation from proteases, wherein the reporter sequence is kkafpaayiae (SEQ ID NO:40);
- a targeting sequence coupled to the reporter sequence and configured to bind to a protein interaction domain of a kinase;
- and
- a transduction sequence coupled to the targeting sequence and configured to resist degradation from proteases, wherein said transduction sequence comprises an amino acid sequence of D-amino acids.
A method of quantifying phosphorylation of a kinase comprising:
- providing a sample including at least one cell having a kinase of interest;
- contacting the sample with a biosensor comprising a reporter sequence configured to resist degradation from proteases, a targeting sequence coupled to the reporter sequence and configured to bind to a protein interaction domain of a kinase, and a transduction sequence coupled to the targeting sequence and configured to resist degradation from proteases, wherein the reporter sequence is kkafpaayiae (SEQ ID NO:40), the transduction sequence comprises an amino acid sequence of D-amino acids, and the biosensor includes a cleavable linker coupled between the reporter sequence and the targeting sequence;
- cleaving the cleavable linker on the biosensor;
- collecting the reporter sequence;
- and quantifying phosphorylation of the sample.
Dependencies are inferred from the claim text.