Multivalent photoaffinity probe for labeling small molecule binding proteins.
Characterization of small molecule (SM)-protein interaction is of high importance in biomedical research such as target identification and proteomic profiling. Photo-cross-linking is a powerful and straightforward strategy to covalently capture SM's binding proteins. The DNA-based photoaffinity labeling method is able to capture SM's protein targets with high specificity but suffers low cross-linking efficiency, which limits its utility for low abundance and low affinity proteins. After screening a variety of cross-linkers, by utilizing the multivalency effect, the cross-linking efficiency was improved by nearly 7-fold without compromising probe specificity. The generality and performance of multivalent photoaffinity probes have been validated with a variety of SM-protein pairs in the complexity of cell lysates.
Key Laboratory of Bioorganic Chemistry and Molecular Engineering of the Ministry of Education, Beijing National Laboratory of Molecular Sciences (BNLMS), College of Chemistry and Molecular Engineering, Peking University , Beijing, China 100871.
Gang
G
Li
Yu
Y
Liu
Xuerong
X
Yu
Xiaoyu
X
Li
D000818:Animals; D001665:Binding Sites; D024402:Carbonic Anhydrase II / Q000032:analysis* / Q000378:metabolism; D002417:Cattle; D003432:Cross-Linking Reagents / Q000138:chemical synthesis / Q000737:chemistry*; D004247:DNA / Q000138:chemical synthesis / Q000737:chemistry; D005453:Fluorescence; D015394:Molecular Structure; D008970:Molecular Weight; D010206:Papain / Q000032:analysis* / Q000378:metabolism; D019920:Photoaffinity Labels / Q000138:chemical synthesis / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000032:analysis*; D022061:Tacrolimus Binding Protein 1A / Q000032:analysis* / Q000378:metabolism
D003432:Cross-Linking Reagents; D019920:Photoaffinity Labels; D012710:Serum Albumin, Bovine; D004247:DNA; D010206:Papain; D024402:Carbonic Anhydrase II; D022061:Tacrolimus Binding Protein 1A