Engineering aldo-keto reductase 1B10 to mimic the distinct 1B15 topology and specificity towards inhibitors and substrates, including retinoids and steroids
Giménez-Dejoz, Joan (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Weber, Susanne (Research Unit Molecular Endocrinology and Metabolism, Helmholtz Zentrum München)
Fernández-Pardo, Álvaro (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Möller, Gabriele (Research Unit Molecular Endocrinology and Metabolism, Helmholtz Zentrum München)
Adamski, Jerzy (Research Unit Molecular Endocrinology and Metabolism, Helmholtz Zentrum München)
Porté, Sergio (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Parés i Casasampera, Xavier (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Farrés, Jaume (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)

Fecha:	2019
Descripción:	9 pàg.
Resumen:	The aldo-keto reductase (AKR)superfamily comprises NAD(P)H-dependent enzymes that catalyze the reduction of a variety of carbonyl compounds. AKRs are classified in families and subfamilies. Humans exhibit three members of the AKR1B subfamily: AKR1B1 (aldose reductase, participates in diabetes complications), AKR1B10 (overexpressed in several cancer types), and the recently described AKR1B15. AKR1B10 and AKR1B15 share 92% sequence identity, as well as the capability of being active towards retinaldehyde. However, AKR1B10 and AKR1B15 exhibit strong differences in substrate specificity and inhibitor selectivity. Remarkably, their substrate-binding sites are the most divergent parts between them. Out of 27 residue substitutions, six are changes to Phe residues in AKR1B15. To investigate the participation of these structural changes, especially the Phe substitutions, in the functional features of each enzyme, we prepared two AKR1B10 mutants. The AKR1B10 m mutant carries a segment of six AKR1B15 residues (299-304, including three Phe residues)in the respective AKR1B10 region. An additional substitution (Val48Phe)was incorporated in the second mutant, AKR1B10mF48. This resulted in structures with smaller and more hydrophobic binding pockets, more similar to that of AKR1B15. In general, the AKR1B10 mutants mirrored well the specific functional features of AKR1B15, i. e. , the different preferences towards the retinaldehyde isomers, the much higher activity with steroids and ketones, and the unique behavior with inhibitors. It can be concluded that the Phe residues of loop C (299-304)contouring the substrate-binding site, in addition to Phe at position 48, strongly contribute to a narrower and more hydrophobic site in AKR1B15, which would account for its functional uniqueness. In addition, we have investigated the AKR1B10 and AKR1B15 activity toward steroids. While AKR1B10 only exhibits residual activity, AKR1B15 is an efficient 17-ketosteroid reductase. Finally, the functional role of AKR1B15 in steroid and retinaldehyde metabolism is discussed.
Ayudas:	Agencia Estatal de Investigación BFU2011-24176 Agencia Estatal de Investigación BIO2016-78057
Derechos:	Tots els drets reservats.
Lengua:	Anglès
Documento:	Article ; recerca ; Versió acceptada per publicar
Materia:	Enzyme kinetics ; Inhibition ; Retinaldehyde ; Site-directed mutagenesis ; Steroids ; SDG 3 - Good Health and Well-being
Publicado en:	Chemico-Biological Interactions, Vol. 307 (2019) , p. 186-194, ISSN 1872-7786

DOI: 10.1016/j.cbi.2019.04.030
PMID: 31028727

31 p, 986.9 KB

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