Patent ID: 6562563
Filing Date: 2003-05-13
Classification: C07K,C12N,G01N

Abstract:
A method for screening for an agent that alters mitochondrial permeability transition (MPT), comprising the steps of:(a) contacting a host cell comprising a mitochondrion with a candidate agent and an inducer of MPT, wherein the host cell comprises (i) a first nucleic acid expression construct, comprising a promoter operably linked to a polynucleotide encoding a mitochondrial permeability transition pore component polypeptide fused to a polynucleotide encoding a first energy transfer molecule or a variant thereof; and (ii) a second nucleic acid expression construct, comprising a promoter operably linked to a polynucleotide encoding a cyclophilin polypeptide fused to a polynucleotide encoding a second energy transfer molecule or a variant thereof, wherein binding of the mitochondrial permeability transition pore component polypeptide to the cyclophilin polypeptide results in detectable energy transfer between the first and second energy transfer molecules; (b) exposing the cell to an excitation energy; (c) detecting a level of energy transfer between the first and second energy transfer molecules; and (d) comparing the level of energy transfer to a first reference level generated in the absence of candidate agent, and therefrom identifying an agent that alters MPT.