title: High-throughput enzymology and combinatorial mutagenesis for mining cytochrome P450 functions.
abstract: BACKGROUND
  High-throughput (HT) characterization of drugs for potential biotransformation and interaction is routine in pharmaceutical industry.
  
  
  OBJECTIVE
  HT approaches were extended to enzyme studies for identifying combinations of structural elements that control substrate specificity.
  
  
  METHODS
  Structure-based and combinatorial mutagenesis have been applied with success to decipher P450 structure-function relationships. The idea is to measure activities on a library of combinatorial variants of similar structure with a large collection of substrates presenting a similar chemical scaffold. This combinatorial approach is then associated to multivariate statistics to relate functional features to structural determinants.
  
  
  CONCLUSIONS
  A method to measure HT kinetics is presented. The proposed statistical approach is illustrated with tri- and tetracyclic substrates and artificial variant enzymes of the CYP1A subfamily.
authors.affiliation: Centre de Génétique Moléculaire, Laboratoire d'Ingénierie des Protéines Membranaires, CNRS UPR2167, Avenue de la terrasse, 91190 Gif-sur-Yvette, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Urban
authors.forename: Gilles
authors.initials: G
authors.lastname: Truan
authors.forename: Denis
authors.initials: D
authors.lastname: Pompon
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001711:Biotransformation; D020650:Combinatorial Chemistry Techniques; D003198:Computer Simulation; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D015233:Models, Statistical; D008969:Molecular Sequence Data; D015394:Molecular Structure; D016297:Mutagenesis, Site-Directed; D004364:Pharmaceutical Preparations / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D004364:Pharmaceutical Preparations; D003577:Cytochrome P-450 Enzyme System; D019363:Cytochrome P-450 CYP1A1