title: High-throughput detection of drugs binding to proteins using desorption electrospray ionization mass spectrometry.
abstract: In this paper, we present a strategy for screening drugs that bind to proteins by combining centrifugal filtration with desorption electrospray ionization mass spectrometry (DESI-MS). Membrane filtration was used to remove any unbound drugs. Then, drug-protein complexes deposited on the DESI substrate were dissociated during the DESI-MS analytical process, and the liberated drugs were measured. To validate the methodology, the screening of a series of drugs against two types of proteins was performed. Three DNA topoisomerase I (Topo I) inhibitors (camptothecin (CPT), hydroxycamptothecin (OHCPT) and 7-ethyl-10-hydroxycamptothecin (SN-38)) were screened against Topo I and the DNA-Topo I complex using DESI-MS. The results indicated that none of the inhibitors bound to Topo I, because the inhibitors had binding affinities only to the DNA-Topo I complex. Among the three drugs that bound to the DNA-Topo I complex, SN-38 had the strongest relative binding affinity, and CPT had the weakest relative binding affinity. The impact of the DESI spray solvent composition on the analysis of drug-protein complex binding was evaluated. Seven alkaloid drugs were also screened against Topo I using DESI-MS. Berberine and palmatine had good binding affinities. A screening of 21 types of drugs was carried out to determine whether the drugs bound to human serum albumin (HSA). The DESI-MS screening process could be achieved within 1.75min. The study provides a method to qualitatively detect compounds that bind to proteins, showing great potential in drug design and screening.
authors: College of Chemistry, Beijing Normal University, Beijing 100875, PR China.
authors: Chenxi
authors: C
authors: Yao
authors: Na
authors: N
authors: Na
authors: Lingyun
authors: L
authors: Huang
authors: Dacheng
authors: D
authors: He
authors: Jin
authors: J
authors: Ouyang
mesh_terms: D002166:Camptothecin / Q000737:chemistry; D004353:Drug Evaluation, Preclinical; D006801:Humans; D004364:Pharmaceutical Preparations / Q000737:chemistry*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D004364:Pharmaceutical Preparations; D011506:Proteins; D002166:Camptothecin
keywords: Camptothecin; DNA topoisomerase I; Desorption electrospray ionization; Drug screening; High-throughput screening; Mass spectrometry