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Sager, C.P.* ; Weber, S. ; Negri, M.* ; Banachowicz, P. ; Möller, G. ; Adamski, J. ; Hartmann, R.W.* ; Marchais-Oberwinkler, S.*

Homology modeling meets site-directed mutagenesis: An ideal combination to elucidate the topology of 17β-HSD2.

J. Steroid Biochem. Mol. Biol. 206:105790 (2021)
DOI
Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.
17β-Hydroxysteroid dehydrogenase type 2 (17β-HSD2) catalyzes the conversion of highly active estrogens and androgens into their less active forms using NAD+ as cofactor. Substrate and cofactor specificities of 17β-HSD2 have been reported and potent 17β-HSD2 inhibitors have been discovered in a ligand-based approach. However, the molecular basis and the amino acids involved in the enzymatic functionality are poorly understood, as no crystal structure of the membrane-associated 17β-HSD2 exists. The functional properties of only few amino acids are known. The lack of topological information impedes structure-based drug design studies and limits the design of biochemical experiments. The aim of this work was the determination of the 17β-HSD2 topology. For this, the first homology model of 17β-HSD2 in complex with NAD+ and 17β-estradiol was built, using a multi-fragment “patchwork” approach. To confirm the quality of the model, fifteen selected amino acids were exchanged one by one using site directed mutagenesis. The mutants’ functional behavior demonstrated that the generated model was of very good quality and allowed the identification of several key amino acids involved in either ligand or internal structure stabilization. The final model is an optimal basis for further experiments like, for example, lead optimization.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Korrespondenzautor
Schlagwörter 17β-hydroxysteroid Dehydrogenase Type 2 ; Enzyme Kinetics ; Enzyme Structure ; Homology Modeling ; Mutagenesis ; Steroid; Human 17-beta-hydroxysteroid-dehydrogenase Type-2; 3-dimensional Structures; Crystal-structures; Protein-structure; Dehydrogenases; Expression; Breast; Recognition; Alignments; Validation
ISSN (print) / ISBN 0960-0760
e-ISSN 0960-0760
Quellenangaben Band: 206, Heft: , Seiten: , Artikelnummer: 105790 Supplement: ,
Verlag Elsevier
Verlagsort The Boulevard, Langford Lane, Kidlington, Oxford Ox5 1gb, England
Nichtpatentliteratur Publikationen
Begutachtungsstatus Peer reviewed
Institut(e) Molekulare Endokrinologie und Metabolismus (MEM)
Institute of Diabetes and Cancer (IDC)
Förderungen Deutsche Forschungsgemeinschaft (DFG), Germany