Method for detecting activity of SIRTs deacetylase based on one-dimensional HNCO nuclear magnetic resonance spectrum and application

Abstract

The invention discloses a method for detecting the activity of SIRTs deacetylase based on a one-dimensional HNCO nuclear magnetic resonance spectrum and application. Comprising the following steps: transferring H3 protein plasmids into escherichia coli, adding the escherichia coli into a culture solution, adding nitrogen 15 labeled ammonia chloride and carbon 13 labeled acetic acid, and culturing to obtain nitrogen 15 and carbon 13 covalently linked H3 protein; adding nitrogen 15 and carbon 13 covalently linked H3 protein into the reaction buffer solution, and incubating to obtain a system 1; detecting the system 1 by using a one-dimensional HNCO nuclear magnetic resonance spectrum to obtain a spectrogram; adding nitrogen 15 and carbon 13 covalently linked H3 protein and SIRTs into the reaction buffer solution, and incubating to obtain a system 2; detecting the system 2 by using a one-dimensional HNCO nuclear magnetic resonance spectrum to obtain a spectrogram; and comparing the two spectrograms to judge the activity of the SIRTs. The invention relates to a method for evaluating the enzyme activity by monitoring the change of SIRTs substrates in real time by using a nuclear magnetic resonance one-dimensional HNCO spectrum.

Description

technical field [0001] The invention relates to the technical field of enzyme activity detection, in particular to a method and application for detecting SIRTs sirtuin activity based on one-dimensional HNCO nuclear magnetic resonance spectroscopy. Background technique [0002] As the fastest-growing disease in the world today, tumor morbidity and fatality rate, its pathogenesis is complex, highly variable, has become one of the top ten causes of human death, is a major disease that affects human health and normal production and life, Inheritance and genetic abnormalities are generally considered to be the cause of tumorigenesis, while studies over the past few decades have demonstrated that epigenetic regulation is also involved in cancer development. [0003] Epigenetic regulation, also known as epigenetics, epigenetic modification or epigenetics, is the study of genetic changes in the activity or function of a gene. The discipline of heritable changes in gene expression. ...

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Problems solved by technology

[0011] In view of the above-mentioned deficiencies in the prior art, the purpose of the present invention is to provide a method and application for detecting SIRTs sirtuin activity based on one-dimensional HNCO nuclear magnetic resonance spectroscopy, aiming to solve the problems of complex and low accuracy of existing detection methods

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