Mass spectrometry method for screening protein with structure and interaction changes caused by drug

Abstract

The invention relates to a mass spectrometry method for rapidly screening a specific protein with structure and interaction changes caused by drug molecules from a complex cell or tissue lysed proteinsample. The mass spectrometry analysis method is specifically an activity-denaturation two-step stable isotope covalent chemical labeling method. In the two-step labeling, an isotope mass differencelabeling reagent is used, and the protein with the structure and interaction being changed by the drug molecules causes changes of the labeling efficiency of a change region in the first step of activity labeling process, so that the difference of isotope peaks is shown in a final mass spectrum quantitative result. The method provides a brand-new high-throughput mass spectrometry method for rapidly screening a target protein with structure and interaction changes caused by drug molecule binding in a complex system.

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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