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Inducible reconstitution and real-time quantitative kinetic system for the study of intramembrane enzymes
Inducible reconstitution and real-time quantitative kinetic system for the study of intramembrane enzymes
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机译:诱导重构和实时定量动力学系统用于膜内酶的研究
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摘要
The present invention relates to the field of enzymes. More specifically, the present invention provides an inducible reconstitution and real-time quantitative kinetic system for the study of intramembrane enzymes. In a specific embodiment, a method of screening for modulators of an intramembrane protease comprises the steps of (a) contacting in a mixture the protease and a substrate with a lipid under acidic or basic conditions to form a membrane comprising the lipid bilayer, protease and the substrate; (b) contacting a test agent with the membrane mixture; (c) adjusting the pH to physiological conditions; (d) assaying substrate cleavage by the protease; and (e) comparing the assayed substrate cleavage to a reference that does not include the test agent, wherein an increase or a decrease of substrate cleavage by the protease relative to the reference identifies the test agent as a modulator of the intramembrane protease.
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