title: Examining the interactome of huperzine A by magnetic biopanning.
abstract: Huperzine A is a bioactive compound derived from traditional Chinese medicine plant Qian Ceng Ta (Huperzia serrata), and was found to have multiple neuroprotective effects. In addition to being a potent acetylcholinesterase inhibitor, it was thought to act through other mechanisms such as antioxidation, antiapoptosis, etc. However, the molecular targets involved with these mechanisms were not identified. In this study, we attempted to exam the interactome of Huperzine A using a cDNA phage display library and also mammalian brain tissue extracts. The drugs were chemically linked on the surface of magnetic particles and the interactive phages or proteins were collected and analyzed. Among the various cDNA expressing phages selected, one was identified to encode the mitochondria NADH dehydrogenase subunit 1. Specific bindings between the drug and the target phages and target proteins were confirmed. Another enriched phage clone was identified as mitochondria ATP synthase, which was also panned out from the proteome of mouse brain tissue lysate. These data indicated the possible involvement of mitochondrial respiratory chain matrix enzymes in Huperzine A's pharmacological effects. Such involvement had been suggested by previous studies based on enzyme activity changes. Our data supported the new mechanism. Overall we demonstrated the feasibility of using magnetic biopanning as a simple and viable method for investigating the complex molecular mechanisms of bioactive molecules.
authors.affiliation: School of Pharmacy, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
authors.forename: Wei
authors.initials: W
authors.lastname: Guo
authors.forename: Shupeng
authors.initials: S
authors.lastname: Liu
authors.forename: Jinliang
authors.initials: J
authors.lastname: Peng
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Wei
authors.forename: Ye
authors.initials: Y
authors.lastname: Sun
authors.forename: Yangsheng
authors.initials: Y
authors.lastname: Qiu
authors.forename: Guangwei
authors.initials: G
authors.lastname: Gao
authors.forename: Peng
authors.initials: P
authors.lastname: Wang
authors.forename: Yuhong
authors.initials: Y
authors.lastname: Xu
mesh_terms: D000470:Alkaloids / Q000494:pharmacology*; D000818:Animals; D001435:Bacteriophages / Q000187:drug effects / Q000378:metabolism; D001921:Brain / Q000187:drug effects* / Q000378:metabolism; D004579:Electron Transport / Q000187:drug effects; D015723:Gene Library; D008280:Magnetics / Q000379:methods*; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D008516:Medicine, Chinese Traditional; D051379:Mice; D008810:Mice, Inbred C57BL; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D025261:Mitochondrial Proton-Translocating ATPases / Q000378:metabolism; D009245:NADH Dehydrogenase / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology*; D010936:Plant Extracts / Q000494:pharmacology*; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism; D020543:Proteome / Q000187:drug effects / Q000378:metabolism; D012717:Sesquiterpenes / Q000494:pharmacology*
chemical_list: D000470:Alkaloids; D058185:Magnetite Nanoparticles; D018696:Neuroprotective Agents; D010936:Plant Extracts; D011506:Proteins; D020543:Proteome; D012717:Sesquiterpenes; C050426:huperzine A; D009245:NADH Dehydrogenase; D025261:Mitochondrial Proton-Translocating ATPases