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Cited by 92Enzyme Structure and FunctionAldose Reductase and TaurineProtein Structure and Dynamics

Structure of 3α-Hydroxysteroid/Dihydrodiol Dehydrogenase Complexed with NADP+

Bennett, Melanie J., Schlegel, Brian P., Jez, Joseph M., Penning, Trevor M., Lewis, Mitchell·1996·Biochemistry
DOI 10.1021/bi9604688
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Abstract

Rat liver 3 alpha-hydroxysteroid/dihydrodiol dehydrogenase (3 alpha-HSD) inactivates circulating steroid hormones and is involved in polycyclic aromatic hydrocarbon (PAH) carcinogenesis. It is the only HSD of known structure in the aldo-keto reductase (AKR) superfamily and may provide a paradigm for other mammalian HSDs in this family. The structure of the 3 alpha-HSD.NADP+ binary complex has been determined at 2.7 A resolution and refined to a crystallographic R-factor of 23.4% with good geometry. The model is similar to other binary complexes in the AKR superfamily in that NADP+ binds at the C-terminal end of an alpha/beta barrel. However, it is unique in that NADP+ is bound in two alternate conformations, probably because of the lack of a salt-linked "safety belt" over the pyrophosphate bridge. The structure supports a previously proposed catalytic mechanism for carbonyl reduction in which Tyr 55 is the general acid, and its effective pKa is lowered by the adjacent Lys 84. We present evidence that the structurally distinct short-chain dehydrogenase/reductase (SDR) superfamily may have convergently evolved a similar catalytic mechanism. Insight into substrate binding is offered by a crystal packing contact in which a neighboring molecule inserts a tryptophan residue (Trp 227) into an apolar cleft in 3 alpha-HSD. This cleft is proximal to the bound NADP+ cofactor and contains a surface of apolar residues (Leu 54, Trp 86, Leu 122, Phe 128, Phe 129, Leu 137, Phe 139), making it a likely candidate for the substrate-binding site. Thus, in forming this crystal contact, Trp 227 may mimic a portion of a bound steroid. In addition, we propose that a water molecule in the active site indicates the position of the hydroxyl oxygen in a 3 alpha-hydroxysteroid substrate. Knowledge of the position of this water molecule, combined with the stereochemistry of hydride transfer, suggests that the alpha face of a bound steroid will be oriented toward the side of the apolar cleft containing Trp 86.

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Crossref · 10.1021/bi9604688

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  • HSDs from the SDR and AKR superfamilies differ in protein fold, hydride-transfer stereochemistry and kinetic mechanism but share a conserved catalytic mechanism.

    “HSDs from the SDR and AKR superfamilies differ in their protein folds, stereochemistry of hydride transfer and kinetic mechanism but share a common catalytic mechanism that has been evolutionary conserved”

    From Role of aldo-keto reductase family 1 (AKR1) enzymes in human steroid metabolism (2014) · cites this paper as [9]

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Authors
Bennett, M. J., Schlegel, B. P., Jez, J. M., Penning, T. M., & Lewis, M.
Affiliations
University of Pennsylvania (US)
ORCID
Venue
Biochemistry
Publisher
American Chemical Society (ACS)
Published
1996-01-01
Volume
35
Issue
33
Pages
10702-10711
ISSN
  • 0006-2960 (ISSN-L)
  • 1520-4995
DOI
10.1021/bi9604688
OpenAlex
W1978718013
Cited by
92
References
19
Citations by year
2024: 1 · 2023: 1 · 2022: 2 · 2021: 3 · 2020: 1 · 2018: 1 · 2017: 2 · 2016: 2 · 2014: 2 · 2013: 1 · 2012: 2
FWCI
7.09
Type
journal article
Language
en
Open access
closed
Locations
  • Biochemistry (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Institutes of Health (5r01ca055711-07)
  • Office of Naval Research
  • National Cancer Institute (CA 39504, CA-55711, R01CA039504)
  • National Institute of Diabetes and Digestive and Kidney Diseases (DK-47015, R01DK047015)
Topics
  • Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Aldose Reductase and Taurine
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
Keywords
NADP+ binding, aldo-keto reductase superfamily, short-chain dehydrogenase/reductase superfamily, steroid hydroxylation, catalytic mechanism, substrate binding site
MeSH terms
Animals, Binding Sites, Catalysis, Liver, Male, Models, Molecular, NADP, Steroids, 3-Hydroxysteroid Dehydrogenases, Molecular Structure, Rats, Sprague-Dawley, Crystallography, X-Ray, Rats

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