title: Substrate Specificity, Inhibitor Selectivity and Structure-Function Relationships of Aldo-Keto Reductase 1B15: A Novel Human Retinaldehyde Reductase.
abstract: Human aldo-keto reductase 1B15 (AKR1B15) is a newly discovered enzyme which shares 92% amino acid sequence identity with AKR1B10. While AKR1B10 is a well characterized enzyme with high retinaldehyde reductase activity, involved in the development of several cancer types, the enzymatic activity and physiological role of AKR1B15 are still poorly known. Here, the purified recombinant enzyme has been subjected to substrate specificity characterization, kinetic analysis and inhibitor screening, combined with structural modeling. AKR1B15 is active towards a variety of carbonyl substrates, including retinoids, with lower kcat and Km values than AKR1B10. In contrast to AKR1B10, which strongly prefers all-trans-retinaldehyde, AKR1B15 exhibits superior catalytic efficiency with 9-cis-retinaldehyde, the best substrate found for this enzyme. With ketone and dicarbonyl substrates, AKR1B15 also shows higher catalytic activity than AKR1B10. Several typical AKR inhibitors do not significantly affect AKR1B15 activity. Amino acid substitutions clustered in loops A and C result in a smaller, more hydrophobic and more rigid active site in AKR1B15 compared with the AKR1B10 pocket, consistent with distinct substrate specificity and narrower inhibitor selectivity for AKR1B15.
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Biosciences, Universitat Autònoma de Barcelona, Bellaterra, Barcelona, Spain.
authors.forename: Joan
authors.initials: J
authors.lastname: Giménez-Dejoz
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic; Institute of Neuroscience and Medicine (INM-9) and Institute for Advanced Simulation (IAS-5), Forschungszentrum Jülich GmbH, Jülich, Germany.
authors.forename: Michal H
authors.initials: MH
authors.lastname: Kolář
authors.affiliation: Department of Integrative Structural Biology, Institut de Génétique et de Biologie Moléculaire et Cellulaire-Centre de Biologie Intégrative, CNRS, INSERM, UdS, Illkirch CEDEX, France.
authors.forename: Francesc X
authors.initials: FX
authors.lastname: Ruiz
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Biosciences, Universitat Autònoma de Barcelona, Bellaterra, Barcelona, Spain.
authors.forename: Isidro
authors.initials: I
authors.lastname: Crespo
authors.affiliation: Department of Integrative Structural Biology, Institut de Génétique et de Biologie Moléculaire et Cellulaire-Centre de Biologie Intégrative, CNRS, INSERM, UdS, Illkirch CEDEX, France.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Cousido-Siah
authors.affiliation: Department of Integrative Structural Biology, Institut de Génétique et de Biologie Moléculaire et Cellulaire-Centre de Biologie Intégrative, CNRS, INSERM, UdS, Illkirch CEDEX, France.
authors.forename: Alberto
authors.initials: A
authors.lastname: Podjarny
authors.affiliation: Diabetes and Obesity Center, School of Medicine, University of Louisville, Louisville, Kentucky, United States of America.
authors.forename: Oleg A
authors.initials: OA
authors.lastname: Barski
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
authors.forename: Jindřich
authors.initials: J
authors.lastname: Fanfrlík
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Biosciences, Universitat Autònoma de Barcelona, Bellaterra, Barcelona, Spain.
authors.forename: Xavier
authors.initials: X
authors.lastname: Parés
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Biosciences, Universitat Autònoma de Barcelona, Bellaterra, Barcelona, Spain.
authors.forename: Jaume
authors.initials: J
authors.lastname: Farrés
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Biosciences, Universitat Autònoma de Barcelona, Bellaterra, Barcelona, Spain.
authors.forename: Sergio
authors.initials: S
authors.lastname: Porté
mesh_terms: D000449:Aldehyde Reductase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D000074408:Aldo-Keto Reductases; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D020134:Catalytic Domain / Q000235:genetics; D004224:Diterpenes; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D050837:Oxidoreductases Acting on Aldehyde or Oxo Group Donors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012172:Retinaldehyde / Q000378:metabolism*; D040681:Structural Homology, Protein; D013379:Substrate Specificity
chemical_list: D004224:Diterpenes; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; C031390:9-cis-retinal; C120471:AKR1B10 protein, human; C560504:AKR1B15 protein, human; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase; D050837:Oxidoreductases Acting on Aldehyde or Oxo Group Donors; D012172:Retinaldehyde