A mass spectrometric approach to screening proteins for drug-induced structural and interaction changes

Abstract

The patent of the present invention relates to a mass spectrometry analysis method for rapidly screening specific proteins whose structures and interactions are changed by drug molecules from complex cell or tissue lysed protein samples. Specifically, it is an "active-denatured two-step stable isotope covalent chemical labeling method". In the two-step labeling, the isotope mass difference labeling reagent is used, and the protein whose structure and interaction are changed by the small drug molecule is actively labeled in the first step. The process will cause changes in the labeling efficiency of the changing regions, thus showing differences in isotope peaks in the final mass spectrometry results. This method provides a new high-throughput mass spectrometry method for rapid screening of target proteins whose structures and interactions change due to the binding of drug molecules in complex systems.

Description

technical field [0001] The invention belongs to the field of mass spectrometry analysis method for quickly screening proteins with changes in structure and interaction caused by drugs in complex systems, in particular to a method for quickly screening specific proteins with changes in structure and interaction caused by drug molecules from complex cell or tissue lysed protein samples new method of mass spectrometry. Background technique [0002] As the executor of life activities, protein almost participates in and controls all life activities in organisms. The combination of drugs and proteins will cause changes in the protein structure, how to effectively detect the interaction between drugs and proteins plays an important role in biochemical research and drug development. In recent years, the analysis method of drug-protein interaction based on mass spectrometry has been gradually developed. Compared with other techniques such as X-ray crystallography and nuclear magnet...

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Problems solved by technology

But these methods usually need more stringent conditions, such as restriction proteolysis technology needs to strictly control enzymolysis conditions (document 2.Piazza I; Kochanowski K; Cappelletti V; et al.A Map of Protein-Metabolite InteractionsReveals Principles of Chemical Communication .Cell 2018, 172(1-2), 358-372), cross-linked mass spectrometry faces difficulties in data processing (document 3.Ferber M, Kosinski J, Ori A, et al.Automated structure modeling of large protein assemblies using crosslinks as distancerestraints.Nature methods,2016,13(6):515-520), the chemical proteomics technology based on active molecular probes needs to design and synthesize probes to make them accurately reach the protein binding region (document 4. Bachovchin D A, Brown S J, Rosen H, et al. Identification of selective inhibitors of uncharacterized enzymes by high-throughput screening with fluorescent activity-based probes. Nature biotechnology, 2009, 27 (4): 387-394)

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