Patent ID: 8999896
Filing Date: 2015-04-07
Classification: C12Q,G01N

Abstract:
1. A method of identifying an agent that modulates N α -terminal acetyl transferase-mediated N-end rule pathway-mediated acetylation of an amino terminal (N-terminal) Met, Ala, Val, Ser, Thr, Cys, Gly, or Pro residue of a peptide, comprising: a) contacting at least one sample comprising a peptide having an N-terminal Met, Ala, Val, Ser, Thr, Cys, Gly, or Pro residue of the peptide, wherein the peptide comprises an N-terminal portion of a fusion protein and wherein the fusion protein comprises a reporter polypeptide C-terminal to the peptide, with at least one test agent, under conditions suitable for the N-terminal Met, Ala, Val, Ser, Thr, Cys, Gly, or Pro residue of the peptide to act as a substrate for an N-end rule pathway reaction; b) using mass spectroscopy or capillary electrophoresis, analyzing the results to detect a change in the N-end rule pathway substrate activity of the N-terminal Met, Ala, Val, Ser, Thr, Cys, Gly, or Pro residue of the peptide of a) in the presence of the test agent as compared to the activity in the absence of the test agent; wherein a change identifies the test agent as an agent that modulates N