Site-Selective Protein Chemical Modification of Exposed Tyrosine Residues Using Tyrosine Click Reaction

• Published in: Bioconjugate chemistry Vol. 31; no. 5; pp. 1417 - 1424
• Main Authors: Sato, Shinichi, Matsumura, Masaki, Kadonosono, Tetsuya, Abe, Satoshi, Ueno, Takafumi, Ueda, Hiroshi, Nakamura, Hiroyuki
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 20-05-2020
• Subjects: Amino acids; Animals; Antibodies; Aromatic compounds; Binding Sites; Biocatalysis; Cell Line, Tumor; Cell Transformation, Neoplastic; Chemical modification; Chemical reactions; Click Chemistry; Complementarity-determining region; Electrochemistry; Electron transfer; Electron Transport; Exposure; Horseradish peroxidase; Horseradish Peroxidase - metabolism; Humans; Hydrophobicity; Luminol - chemistry; Male; Methylation; Mice; Models, Molecular; Peroxidase; Protein Conformation; Proteins; Proteins - chemistry; Residues; Single electrons; Tyrosine; Tyrosine - chemistry
• ISSN: 1043-1802; 1520-4812
• DOI: 10.1021/acs.bioconjchem.0c00120

Targeting less abundant amino acid residues on the protein surface may realize site-selective protein modification of natural proteins. The relative hydrophobicity of tyrosine combined with the π–π stacking tendency of the aromatic rings results in generally low accessibility. In this study, site-selective protein modification was achieved by targeting surface-exposed tyrosine residues without using a genetic encoding system. Tyrosine residues were modified with N-methylated luminol derivative under single-electron transfer (SET) reaction conditions. Horseradish peroxidase (HRP)-catalyzed SET and electrochemically activated SET modified surface-exposed tyrosine residues selectively. N-Methylated luminol derivative modified tyrosine residues more efficiently than 4-arylurazole under tyrosine click conditions using HRP and electrochemistry. Tyrosine residues that are evolutionarily exposed only in the complementarity-determining region (CDR) of an antibody were selectively modified by tyrosine click reactions. CDR-modified antibodies were applied to in vivo imaging and antibody–drug conjugated (ADC).