title: Stereoselective binding of mexiletine and ketoprofen enantiomers with human serum albumin domains.
abstract: OBJECTIVE
  To investigate the stereoselective binding of mexiletine or ketoprofen enantiomers with different recombinant domains of human serum albumin (HSA).
  
  
  METHODS
  Three domains (HSA DOM I, II and III) were expressed in Pichia pastoris GS115 cells. Blue Sepharose 6 Fast Flow was employed to purify the recombinant HSA domains. The binding properties of the standard ligands, digitoxin, phenylbutazone and diazepam, and the chiral drugs to HSA domains were investigated using ultrafiltration. The concentrations of the standard ligands, ketoprofen and mexiletine were analyzed with HPLC.
  
  
  RESULTS
  The recombinant HSA domains were highly purified as shown by SDS-PAGE and Western blotting analyses. The standard HSA ligands digitoxin, phenylbutazone and diazepam selectively binds to DOM I, DOM II and DOM III, respectively. For the chiral drugs, R-ketoprofen showed a higher binding affinity toward DOM III than S-ketoprofen, whereas S-mexiletine bound to DOM II with a greater affinity than R-mexiletine.
  
  
  CONCLUSIONS
  The results demonstrate that HSA DOM III possesses the chiral recognition ability for the ketoprofen enantiomers, whereas HSA DOM II possesses that for the mexiletine enantiomers.
authors.affiliation: Department of Drug Metabolism and Pharmaceutical Analysis, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China.
authors.forename: Da
authors.initials: D
authors.lastname: Shi
authors.forename: Yin-xiu
authors.initials: YX
authors.lastname: Jin
authors.forename: Yi-hong
authors.initials: YH
authors.lastname: Tang
authors.forename: Hai-hong
authors.initials: HH
authors.lastname: Hu
authors.forename: Si-yun
authors.initials: SY
authors.lastname: Xu
authors.forename: Lu-shan
authors.initials: LS
authors.lastname: Yu
authors.forename: Hui-di
authors.initials: HD
authors.lastname: Jiang
authors.forename: Su
authors.initials: S
authors.lastname: Zeng
mesh_terms: D001665:Binding Sites; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007536:Isomerism; D007660:Ketoprofen / Q000737:chemistry / Q000378:metabolism*; D008024:Ligands; D008801:Mexiletine / Q000737:chemistry / Q000378:metabolism*; D015394:Molecular Structure; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D012709:Serum Albumin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship; D014462:Ultrafiltration
chemical_list: D008024:Ligands; D011994:Recombinant Proteins; D012709:Serum Albumin; D008801:Mexiletine; D007660:Ketoprofen