title: Immunoprecipitation coupled with HPLC-MS/MS to discover the aromatase ligands from Glycyrrhiza uralensis.
abstract: Affinity chromatography, applied to discover the enzyme inhibitors, needs special column with target protein and its carrier. Selection of stationary phase and mobile phase needs careful considerations due to the characteristics of proteins. In this study, a method immunoprecipitation (IP) coupled with HPLC-DAD-MS was developed to discover the aromatase ligands from Glycyrrhiza uralensis. An SB-C18 column was employed to separate target compounds without special consideration in mobile phase. Twenty-one compounds, including isolated compounds 4, 7, 8, 10, 11, 13, 15, 18-20, 23 and non-isolated compounds A-J, were found to have good affinity to aromatase by LC-MS. Seven of them (7, 15, 18, 19, 23, D, E) were detected to bind with aromatase in MCF-7 cells by IP coupled with HPLC-MS/MS. Bioassays disclosed aromatase inhibitory activities of the isolated compounds mentioned above, verifying the efficiency of IP coupled with HPLC-MS/MS as a method to screen aromatase ligands.
authors.affiliation: Second Affiliated Hospital Zhejiang University School of Medicine, No. 88 Jiefang Rd., Hangzhou 310009, China.
authors.forename: Liping
authors.initials: L
authors.lastname: Luo
authors.forename: Liming
authors.initials: L
authors.lastname: Shen
authors.forename: Fang
authors.initials: F
authors.lastname: Sun
authors.forename: Zhongjun
authors.initials: Z
authors.lastname: Ma
mesh_terms: D001141:Aromatase / Q000737:chemistry; D047072:Aromatase Inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D029882:Glycyrrhiza uralensis / Q000737:chemistry*; D006801:Humans; D047468:Immunoprecipitation; D007700:Kinetics; D008024:Ligands; D010936:Plant Extracts / Q000737:chemistry*; D011485:Protein Binding; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D047072:Aromatase Inhibitors; D004791:Enzyme Inhibitors; D008024:Ligands; D010936:Plant Extracts; D001141:Aromatase