Direct observation of molecular modifications in biological test systems by measuring flourescence lifetime
The invention relates to a method for directly detecting the modification of a molecule containing a fluorescent dye by measuring the fluorescence lifetime.
1 . A method of homogeneously, directly and quantitatively measuring molecule modifications, characterized in that the molecule carries a fluorescent dye and that the fluorescence lifetime of said molecule differs from the fluorescence lifetime of the modified molecule.
2 . The method of claim 1 , in which the molecule is an organic molecule, or is an inorganic molecule.
3 . The method of claim 1 , wherein the fluorescent dye is a coumarine, a fluoresceine, a rhodarnine, an oxazine, or a cyanine dye.
4 . The method of claim 1 , wherein the fluorescent dye is covalently or noncovalently coupled to the molecule and optionally a spacer molecule may be located between the fluorescent dye and the molecule.
5 . The method of claim 1 for quantifying biochemical assays.
6 . The method of claim 5 , wherein enzymes can carry out the following modification reactions:
phosphorylation/dephosphorylation, sulfation/desulfation, methylation/demethylation, oxidations/reductions, acetylation/deacetylation, amidation/deamidation, cyclization/decyclization, conformational changes, removal of amino acids/peptides/coupling of amino acids/peptides, ring expansion/ring contraction, rearrangements, substitutions, eliminations, addition reactions.
7 . The method of claim 1 for the use in high throughput screening.
8 . A reagent kit comprising fluorescent dye-molecule conjugates and other reagents required for carrying out the assay method as claimed in claims 1 to 6 .
9 . The method of claim 2 , wherein the organic molecule is a peptide or peptidomimetic.