Method of measuring the activity of lipid-modified enzyme
AssigneeAjinomoto Co., Inc.
InventorNaoyuki FUKUCHI
Life of this patent
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Abstract
It is intended to provide an assay method whereby the activity of a lipid-modifying enzyme can be conveniently measured over a wide range and a drug capable of controlling a lipid-modifying enzyme with the use of this assay method. The above problem can be solved by, for example, a method of measuring the activity of a lipid-modifying enzyme which comprises the steps of (I) preparing a lipid micelle containing a biotinylated lipid and a substrate for the lipid-modifying enzyme; (II) bringing the lipid micelle prepared in the above step (I) into contact with the lipid-modifying enzyme; and (III) evaluating the activity of the lipid-modifying enzyme by applying an evaluation method using the proximity effect to the product obtained in the above step (II).
Claims16
A method of assaying the activity of a lipid-modifying enzyme, wherein the method comprises:
- bringing a lipid micelle comprising a biotinylated lipid and a substrate for the lipid-modifying enzyme into contact with the lipid-modifying enzyme to obtain a product;
- and
- evaluating the activity of the lipid-modifying enzyme by applying an evaluation method using a proximity effect to the product obtained from said bringing.
A method of screening a substance inhibiting or promoting the activity of a lipid-modifying enzyme, wherein the method comprises:
- bringing a lipid micelle comprising a biotinylated lipid and a substrate for the lipid-modifying enzyme into contact with the lipid-modifying enzyme in the presence or absence of an analyte to obtain a product;
- evaluating the activity of the lipid-modifying enzyme by applying an evaluation method using a proximity effect to the product obtained from said bringing;
- and
- comparing the activity of the lipid-modifying enzyme in the presence of an analyte with the activity of the lipid-modifying enzyme in the absence of the analyte to determine that the analyte is a substance inhibiting the activity of the lipid-modifying enzyme or a substance promoting the activity of the lipid-modifying enzyme.
A method of screening a substance inhibiting or promoting the activity of a lipid-modifying enzyme, wherein the method comprises:
- bringing a lipid micelle comprising a biotinylated lipid and a substrate for the lipid-modifying enzyme into contact with the lipid-modifying enzyme in the presence of an analyte, and in the presence or absence of a lipid membrane-bound protein having a controlling function to the activity of the lipid-modifying enzyme to be analyzed to obtain a product;
- evaluating the activity of the lipid-modifying enzyme by applying an evaluation method using a proximity effect to the product obtained from said bringing;
- and
- comparing the activity of the lipid-modifying enzyme in the presence of the lipid membrane-bound protein with the activity of the lipid-modifying enzyme in the absence of the lipid membrane-bound protein to determine that the analyte is a substance inhibiting the activity of the lipid-modifying enzyme or a substance promoting the activity of the lipid-modifying enzyme through affecting interactions between the lipid membrane-bound protein and the lipid-modifying enzyme.
Dependencies are inferred from the claim text.