Methods for transferases
OwnerSaid Goueli, Robert Bulleitinventors
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Abstract
Methods for detecting transferase activity of a sample are provided. Transferase activity detected includes kinase and phosphatase activity. The method includes providing a substrate having a reporter compound conjugated thereto. A quantity of the substrate is added to a solution containing the sample. The sample is incubated with the substrate under conditions where the sample is active for a time sufficient for kinase or phosphatase activity to take place. An exopeptidase is added to the solution containing the sample. Output of the reporter compound is detected.
Claims18
A method for detecting kinase or phosphatase activity of a sample, the method comprising:
- (A) contacting the sample with a peptide substrate and at least one of a phosphate group donor and a phosphate group acceptor, wherein the substrate includes a reporter compound and amino acids;
- (B) adding an exopeptidase, wherein phosphorylation of the substrate affects cleavage by the exopeptidase, wherein adding the exopeptidase comprises adding an exopeptidase that hydrolyzes the peptide substrate from a carboxy-terminal end of the substrate;
- and
- (C) detecting output of the reporter compound, wherein the reporter compound exhibits a different output property when bound to at least one amino acid of the substrate when compared to when it is not bound to amino acids.
A method of detecting kinase or phosphatase activity of a sample comprising:
- (A) providing a substrate having a reporter compound conjugated thereto;
- (B) adding a quantity of the substrate to a solution containing the sample;
- (C) incubating the sample with the substrate under conditions where the sample is active for a time sufficient for kinase or phosphatase activity to take place;
- (D) adding an exopeptidase to the solution containing the sample;
- (E) detecting output of the reporter compound.
A method for detecting peptide transferase activity of a sample, the method comprising:
- (A) contacting the sample with a peptide substrate and at least one of a group donor and a group acceptor, wherein the substrate includes a reporter compound and amino acids;
- (B) adding an exopeptidase, wherein transfer of a group from one substrate to another affects cleavage of the substrate by the exopeptidase;
- and
- (C) detecting output of the reporter compound.
A method for detecting peptide transferase activity of a sample, the method comprising:
- (A) contacting the sample with a peptide substrate and at least one of a group donor and a group acceptor, wherein the substrate includes a reporter compound and two peptides, each peptide being separately linked to the reporter compound;
- and
- (B) adding an exopeptidase, wherein transfer of a group from one substrate to another affects cleavage of the substrate by the exopeptidase;
- and
- (C) detecting output of the reporter compound.
A method for detecting peptide transferase activity of a sample, the method comprising:
- (A) contacting the sample with a peptide substrate and at least one of a group donor and a group acceptor, wherein the substrate includes a reporter compound and amino acids;
- and
- (B) adding an exopeptidase, wherein transfer of a group from one substrate to another affects cleavage of the substrate by the exopeptidase;
- and
- (C) liberating the reporter compound from the substrate.
Dependencies are inferred from the claim text.