Method for identifying modulators of 4-hydroxyphenylpyruvate dioxygenase, useful potentially as herbicides, by fluorescent polarization detection of interaction, also new modulators
Method for identifying substances (I) that interact with 4-hydroxyphenylpyruvate dioxygenase (4-HPD). Method for identifying substances (I) that interact with 4-hydroxyphenylpyruvate dioxygenase (4-HPD) comprises (1) preparing a mixture of 4-HPD, a fluorescent agent (II) that interacts with 4-HPD an...
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Zusammenfassung: | Method for identifying substances (I) that interact with 4-hydroxyphenylpyruvate dioxygenase (4-HPD). Method for identifying substances (I) that interact with 4-hydroxyphenylpyruvate dioxygenase (4-HPD) comprises (1) preparing a mixture of 4-HPD, a fluorescent agent (II) that interacts with 4-HPD and a test compound (or mixture) in different concentrations; (2) irradiating the mixture with plane polarized light that excites emission from (II); and (3) measuring the fluorescent polarization value (FPV) or the anisotropy value of the emitted light. A reduction in FPV with increasing concentration of test compound indicates an interaction with 4-HPD. Independent claims are also included for the following: (1) similar method for testing if a compound interacts with 4-HPD in which a mixture of (I) and (II), at various concentrations, is irradiated and an increase in FPV with decreasing concentration of (II) indicates an interaction; (2) new substances (IIa) that comprise a fluorescent dye attached, through a linker, to a substance that interacts with 4-HPD; (3) test system comprising containers for mixtures of test compounds, device for irradiating the containers with exciting light and device for measuring FPV or anisotropy value of emitted light; and (4) (I) identified by the new method.
Die vorliegende Erfindung betrifft ein Verfahren zum Auffinden von Substanzen, die mit dem Enzym 4-Hydroxy-phenyl-pyruvat-Dioxygenase (4-HPD) wechselwirken, sowie ein Verfahren zum Testen der Wechselwirkung einer Substanz mit 4-HPD. Des Weiteren betrifft die Erfindung Reagenzien für dieses Verfahren, ein entsprechendes Test-System und 4-HPD-Hemmstoffe, die durch die erfindungsgemäßen Verfahren identifiziert werden. |
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