Protein chemical cross-linking agent, preparation method and applications thereof

Abstract

The invention belongs to the technical field of protein and function research thereof, and particularly relates to a protein chemical cross-linking agent, a preparation method and applications thereof, wherein the structure of the protein chemical cross-linking agent is represented by a general formula I. According to the invention, the protein chemical cross-linking agent has the advantages of rapid reaction and no hydrolysis, shows efficient cross-linking activity when being applied to a CXMS technology, and can realize the specific recognition on arginine and lysine; and the results of thecross-linking reaction of the protein chemical cross-linking agent, six mode proteins, ten protein mixtures, multi-subunit yeast protein complex, 70S ribosome and exosome prove that the cross-linkingagent is effectively combined with lysine, so that the problems that the existing cross-linking agents are not rich in type, and the existing succinimide ester cross-linking agents have a lot of problems such as easy hydrolysis, low selectivity, slow cross-linking reaction and the like in the actual process of the CXMS technology are solved.

Description

technical field [0001] The invention belongs to the technical field of protein and its function research, in particular to a protein chemical cross-linking agent and its preparation method and application. Background technique [0002] As the main bearer of life activities, protein plays an important role in the growth and development of organisms. The basic unit of protein is amino acid, and the amino and carboxyl groups of the amino acid backbone are connected to each other to form a polypeptide. The polypeptide is modified by folding to form a protein with a three-dimensional structure. The physiological functions of proteins depend on specific three-dimensional protein structures and protein interaction networks. Therefore, the study of protein spatial structure and protein interaction can further explore the relationship between protein function and structure, and the relationship between protein interactions, and further explain the mystery of life. [0003] Chemical...

AI Technical Summary

Problems solved by technology

[0008] However, due to its easy hydrolysis, low selectivity, and slow cross-linking reaction, succinimide ester cross-linking agents often fail due to hydrolysis during cross-linking experiments, and even if they can be cross-linked, they often get a large number of mono-linked peptide form

The mono-linked peptide form provides limited information for the analysis of protein structure and the understanding of protein interaction, so succinimide ester crosslinkers often need more dosage to obtain enough inter-linked peptide forms, while Adding too much cross-linking agent itself has the risk of destroying the protein structure

[0009] In addition, due to the slow cross-linking reaction, it is difficult to effectively immobilize weak and loose protein complexes, making it difficult for succinimide ester cross-linkers to be used to study weak interactions and structural dynamic changes of proteins

However, the routinely used protein complex fixatives such as formaldehyde and glutaraldehyde are difficult to analyze and apply to CXMS technology due to their poor reaction selectivity.

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