title: [Identification of ligands for human LOX-1 through fluorescence polarization-based high throughput screening].
abstract: OBJECTIVE
  To develop a fluorescence polarization-based high throughput screening and identify ligands for human Lectin-like oxidized low-density lipoprotein receptor-1 (hLOX-1).
  
  
  METHODS
  Sequential ultracentrifugation at 4 degrees C from normolipidemic fasting volunteers to obtain low density lipoprotein (LDL), which was modified by CuSO4 (5 micromol x L(-1)) at 37 degrees C for 24 h. The assay was based on the interaction between receptor and ligand, and hLOX-1 was labeled by FITC and bound to its specific ligand, oxLDL. Different reaction time and DMSO concentration were optimized to determine the stability and tolerance of fluorescence polarization (FP) assay. 3 200 compounds were screened in black 384-well microplate by FP-based competitive displacement assay, at excitation filter of 485 nm and emission filter of 530 nm. Z' was used to assess the assay quality.
  
  
  RESULTS
  The FP-based HTS was formatted in a 384-well microplate with a Z' factor of 0. 75, and three active compounds for hLOX-1 were identified with IC50 below 40 micromol x L(-1) from total 3 200 compounds.
  
  
  CONCLUSIONS
  The results indicated that the fluorescence polarization assay is stable, sensitive, reproducible and well suited for high throughput screening efforts.
authors.affiliation: Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
authors.forename: Tian-Tai
authors.initials: TT
authors.lastname: Zhang
authors.forename: Zhen-Tai
authors.initials: ZT
authors.lastname: Huang
authors.forename: Ying
authors.initials: Y
authors.lastname: Dai
authors.forename: Ai-Lin
authors.initials: AL
authors.lastname: Liu
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
authors.forename: Guan-Hua
authors.initials: GH
authors.lastname: Du
mesh_terms: D001667:Binding, Competitive; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005454:Fluorescence Polarization / Q000379:methods*; D006801:Humans; D008024:Ligands; D008077:Lipoproteins, LDL / Q000378:metabolism*; D051127:Scavenger Receptors, Class E / Q000378:metabolism*
chemical_list: D008024:Ligands; D008077:Lipoproteins, LDL; C495137:OLR1 protein, human; D051127:Scavenger Receptors, Class E; C099252:oxidized low density lipoprotein