Immunochemical and LC–MS/MS Detection of Protein Adduction Resulting from Metabolic Activation of Columbin In Vitro and In Vivo

Columbin (CLB) is a diterpenoid furanolactone compound occurring in some herbal medicines. Administration of CLB has been reported to induce liver injury. The reported CLB hepatotoxicity is suggested to require metabolism to a cis-enedial intermediate. We successfully detected hepatic protein adduct...

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Bibliographic Details
Published in:Chemical research in toxicology 2023-04, Vol.36 (4), p.691-702
Main Authors: Wu, Chutian, Pan, Jie, Gong, Bowen, Liao, Yufen, Zou, Ying, Wang, Yang, Ran, Guangyun, Wang, Xin, Zhou, Mengyue, Liu, Ting, Peng, Ying, Li, Weiwei, Zheng, Jiang
Format: Article
Language:English
Subjects:
Activation, Metabolic
Chromatography, Liquid - methods
Diterpenes - chemistry
Lysine
Proteins - chemistry
Tandem Mass Spectrometry - methods
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Summary:Columbin (CLB) is a diterpenoid furanolactone compound occurring in some herbal medicines. Administration of CLB has been reported to induce liver injury. The reported CLB hepatotoxicity is suggested to require metabolism to a cis-enedial intermediate. We successfully detected hepatic protein adduction resulting from the metabolic activation of CLB and found that the intermediate reacted with lysine residues or lysine/cysteine residues to produce the corresponding pyrroline derivative or pyrrole derivative. The detection was achieved by proteolysis- and liquid chromatography–tandem mass spectrometry (LC–MS/MS)-based methods. Furthermore, we prepared a polyclonal antibody approach which allowed us to detect the protein adduction in the forms of protein immunoblot as well as tissue- and cell-based immunostaining. The antibody technique verified the protein adduction detected by LC–MS/MS.
ISSN:0893-228X
1520-5010
DOI:10.1021/acs.chemrestox.3c00006