title: A Novel Assay for Screening Inhibitors Targeting HIV Integrase LEDGF/p75 Interaction Based on Ni(2+) Coated Magnetic Agarose Beads.
abstract: HIV-1 integrase (IN) plays an essential role in viral replication and thus serves as an important target for chemotherapeutic intervention against HIV-1 infection. However, the current three clinical IN inhibitors, raltegravir, elvitegravir and dolutegravir share the same inhibitory mechanism, resulting in a common clinical resistance profile which have emerged in infected patients receiving treatment. Therefore, it is important to develop small molecule inhibitors that impair IN function with distinct mechanisms of action. In this work, a magnetic-beads based biochemical assay targeting the protein-protein interaction (PPI) between HIV IN and the cellular cofactor LEDGF/p75 was developed for identification of HIV-1 IN inhibitors. Furthermore, a library containing 1000 US. Food and Drug Administration (FDA)-approved drugs currently used for human medication was screened to identify inhibitors targeting the PPI. The assay was proved to be quite robust and with the novel assay we successfully identified dexlansoprazole (IC50 of 4.8 μM), a FDA-approved proton pump inhibitor, as a potential inhibitor for the PPI between IN and LEDGF/p75, which bound to the LEDGF/p75 partner with a kinetic dissociation (Kd) constant of 330 nM ± 2.6 nM.
authors.affiliation: Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing, 100193, China.|Institute of Bioinformatics and Medical Engineering, School of Electrical and Information Engineering, Jiangsu University of Technology, Changzhou, 213001, China.
authors.forename: Zhang
authors.initials: Z
authors.lastname: Dawei
authors.affiliation: Chongqing Center for Biomedicines and Medical Equipment, Chongqing Academy of Science and Technology, Chongqing, 401123, China.
authors.forename: He
authors.initials: H
authors.lastname: Hongqiu
authors.affiliation: Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing, 100193, China.|Institute of Bioinformatics and Medical Engineering, School of Electrical and Information Engineering, Jiangsu University of Technology, Changzhou, 213001, China.
authors.forename: Liu
authors.initials: L
authors.lastname: Mengmeng
authors.affiliation: Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing, 100193, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Zhixia
authors.affiliation: Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing, 100193, China.
authors.forename: Guo
authors.initials: G
authors.lastname: Shunxing
mesh_terms: D001681:Biological Assay / Q000379:methods*; D064748:Dexlansoprazole / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D019427:HIV Integrase / Q000378:metabolism*; D057166:High-Throughput Screening Assays; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry / Q000378:metabolism*; D053759:Interleukin-23 / Q000494:pharmacology; D008280:Magnetics; D008863:Microspheres; D009532:Nickel / Q000737:chemistry*; D010455:Peptides / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D000072417:Protein Domains; D025941:Protein Interaction Mapping; D054328:Proton Pump Inhibitors / Q000737:chemistry / Q000494:pharmacology; D015203:Reproducibility of Results; D012685:Sepharose / Q000737:chemistry*
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D053759:Interleukin-23; D010455:Peptides; D054328:Proton Pump Inhibitors; C119380:lens epithelium-derived growth factor; D009532:Nickel; D012685:Sepharose; D019427:HIV Integrase; D064748:Dexlansoprazole; C514801:p31 integrase protein, Human immunodeficiency virus 1