title: TransCent: computational enzyme design by transferring active sites and considering constraints relevant for catalysis.
abstract: BACKGROUND
  Computational enzyme design is far from being applicable for the general case. Due to computational complexity and limited knowledge of the structure-function interplay, heuristic methods have to be used.
  
  
  RESULTS
  We have developed TransCent, a computational enzyme design method supporting the transfer of active sites from one enzyme to an alternative scaffold. In an optimization process, it balances requirements originating from four constraints. These are 1) protein stability, 2) ligand binding, 3) pKa values of active site residues, and 4) structural features of the active site. Each constraint is handled by an individual software module. Modules processing the first three constraints are based on state-of-the-art concepts, i.e. RosettaDesign, DrugScore, and PROPKA. To account for the fourth constraint, knowledge-based potentials are utilized. The contribution of modules to the performance of TransCent was evaluated by means of a recapitulation test. The redesign of oxidoreductase cytochrome P450 was analyzed in detail. As a first application, we present and discuss models for the transfer of active sites in enzymes sharing the frequently encountered triosephosphate isomerase fold.
  
  
  CONCLUSIONS
  A recapitulation test on native enzymes showed that TransCent proposes active sites that resemble the native enzyme more than those generated by RosettaDesign alone. Additional tests demonstrated that each module contributes to the overall performance in a statistically significant manner.
authors.affiliation: Institut für Biophysik und Physikalische Biochemie, Universität Regensburg, Regensburg, Germany. andre.fischer@biologie.uni-regensburg.de
authors.forename: André
authors.initials: A
authors.lastname: Fischer
authors.forename: Nils
authors.initials: N
authors.lastname: Enkler
authors.forename: Gerd
authors.initials: G
authors.lastname: Neudert
authors.forename: Marco
authors.initials: M
authors.lastname: Bocola
authors.forename: Reinhard
authors.initials: R
authors.lastname: Sterner
authors.forename: Rainer
authors.initials: R
authors.lastname: Merkl
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002384:Catalysis; D020134:Catalytic Domain; D019295:Computational Biology / Q000379:methods*; D030562:Databases, Protein; D004798:Enzymes / Q000737:chemistry*; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D016415:Sequence Alignment; D013329:Structure-Activity Relationship
chemical_list: D004798:Enzymes; D008024:Ligands