The invention provides a polypeptide probe based on photo-crosslinking and nano-
metal particle self-
assembly technologies. The polypeptide probe is modified with
protein-post-translational modified polypeptide. According to the polypeptide probe, affinity recognition and enrichment capture processes are integrated, so that the convenient and efficient recognition and identification of a
protein-post-translational modified
binding protein or a compound of the
binding protein. According to the polypeptide probe of the invention, a photo-crosslinking group is introduced into nano
metal particles,so that a weak acting force between the polypeptide and the
binding protein is converted into a strong
covalent bond effect formed by the photo-crosslinking group, and therefore, the polypeptide probe can bear strong
elution conditions and can capture proteins with low abundance and transient interaction; the influence of the modification of the photo-crosslinking group on a polypeptide frameworkis avoided, and therefore, the original structure and recognition characteristic of the polypeptide framework are well maintained, and influence on the binding specificity of the corresponding binding
protein is greatly reduced. The invention further provides a preparation method of the polypeptide probe, and the application of the polypeptide probe to the identification of post-translational-modified binding proteins, the identification of interaction between protein modifications and the discovery of new protein-modified binding proteins.