Fluorescence Assay For Ghrelin O-Acyltransferase Activity
AssigneeSyracuse University
InventorsJames Hougland, Joseph Darling
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Abstract
An assay to detect ghrelin O-acyltransferase activity using an acrylodan-labeled peptide mimic of ghrelin that provides for high-throughput screening for ghrelin O-acyltransferase inhibitors and detection via high performance liquid chromatography. Alternatively, the assay for ghrelin acylation may be based on a synthetic peptide substrate that mimics the N-terminal sequence of ghrelin and has an environmentally-sensitive fluorophore attached to its C-terminal amino acid through chemoselective ligation.
Claims17
A substrate for use in an assay for ghrelin acylation, comprising a peptide partially corresponding to the N-terminal sequence of human ghrelin O-acyltransferase and including a cysteine mutation to said N-terminal sequence.
An assay for ghrelin acylation, comprising:
- a peptide partially corresponding to the N-terminal sequence of human ghrelin and including a cysteine mutation to said N-terminal sequence;
- and
- a quantity of human ghrelin O-acyltransferase.
A method for detecting whether a target substance inhibits human ghrelin O-acyltransferase, comprising the steps of:
- providing a peptide substrate partially corresponding to the N-terminal sequence of human ghrelin O-acyltransferase and including a cysteine mutation to said N-terminal sequence;
- providing a quantity of human ghrelin O-acyltransferase;
- providing a quantity of said target substance;
- and
- observing any change in production of the octanoylated product peptide.
Dependencies are inferred from the claim text.