Patent ID: 9547007
Date: 2017-01-17
CPC Classifications: G01N

Claim:
1. A method of identifying an enzyme that interacts with a substrate, said method comprising the steps of: a) providing a polypeptide molecule comprising: i) a substrate moiety which interacts with an intracellular enzyme, said substrate moiety comprising a peptide and at least one photoreactive crosslinking moiety; ii) a peptide moiety comprising R1-CPP-R2, wherein CPP is a cell-penetrating peptide, further wherein each of R1 and R2 are independently selected from the group consisting of a peptide, an amino acid, NH2, H, or OH, further wherein said substrate moiety is covalently attached to one of the members selected from the group consisting of R1, R2, a cysteine residue within said peptide moiety, or a lysine (K) residue within said peptide moiety; and iii) a chemical bond linking said substrate moiety and said peptide moiety; b) allowing said molecule to interact with said intracellular enzyme in a cell to form a molecule-enzyme complex, under conditions suitable for binding of said molecule with said enzyme; c) activating said photoreactive crosslinking moiety, thereby covalently cross-linking said substrate moiety with said enzyme that interacts with said substrate; d) isolating said crosslinked substrate-enzyme from said cell; and e) identifying said enzyme crosslinked to said substrate; thereby identifying an enzyme that interacts with a substrate.