METHOD FOR QUANTITATIVE ANALYSIS OF PROLINE HYDROXYLATION-INDUCED INTERACTIONS BETWEEN HIF-1 AND VBC PROTEIN COMPLEX USING FLUORESCENCE POLARIZATION

摘要

A method for analyzing interaction between a HIF-1 peptide and a VBC protein induced by proline hydroxylation is provided to be able to simply analyze the interaction by measuring the change of fluorescence polarization, thereby being effectively applied to an ultra-speed searching using a well-plate. The method comprises the steps of: (a) attaching a fluorescent material to a HIF-1 peptide including a hydroxyproline group to prepare a fluorescent probe; (b) reacting the fluorescent probe with a VBC protein; and (c) after measuring the fluorescence polarization of the reactant, comparing it with the fluorescence polarization of the fluorescent probe itself so as to observe the change of the fluorescence polarization. In the method, the fluorescent probe is prepared by synthesizing a peptide from a specific peptide sequence known to be linked to a VBC protein among amino acid sequences of the known HIF-1 protein, linking an aminocaproic acid linker to an N-terminal of the synthesized peptide, and then labeling a fluorescent material to the end thereof.