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Record W2559744957 · doi:10.1107/s2053273314084964

The trans-membrane 17βHSD7: Kinetic study and preliminary crystallization data

2014· article· en· W2559744957 on OpenAlexaff
Jean-Francois Theriault, Dao‐Wei Zhu, Sheng‐Xiang Lin

Bibliographic record

VenueActa Crystallographica Section A Foundations and Advances · 2014
Typearticle
Languageen
FieldMedicine
TopicHormonal Regulation and Hypertension
Canadian institutionsCentre hospitalier universitaire de Québec
Fundersnot available
KeywordsChemistryCrystallizationEnzyme kineticsEnzymeKineticsCofactorDehydrogenaseSteady state (chemistry)Transmembrane proteinCholesterol oxidaseChromatographyBiochemistryActive siteOrganic chemistry

Abstract

fetched live from OpenAlex

Human 17β-Hydroxysteroid dehydrogenase type 7 (h17HSD7) is a transmembrane enzyme belonging to the large, phylogenetically related protein family of short-chain dehydrogenases/reductases (SDR). In this study, h17HSD7 was expressed, purified, and the steady-state kinetics was studied. Here we report a purification method of the transmembrane enzyme using FPLC equipment with a combination of detergents to obtain homogeneous protein. We also report steady-state kinetics for h17HSD7 at 37 degrees Celsius and pH 7.5, using a homogeneous enzyme preparation with estrone (E1), or dihydrotestosterone (DHT) as substrate and NADPH as the cofactor. Kinetic studies made over a wide range of concentrations of both steroids (2microM to 60microM) revealed that E1 (Km= 12microM) has a higher apparent affinity for the h17BHSD7 compared to DHT (Km= 44microM). Crystallization of h17HSD7 in complex with high concentration of steroids to saturate the enzyme in the drop as low apparent affinity of both steroids was found in kinetic study. Some preliminary crystals were obtained by hanging-drop and diffracted at synchrotron facility (Maximum resolution: 2.95Å , Space group: H 3 2, Average unit cell: 120.74 120.74 201.94 90 90 90, Mosaicity : 0.51).

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.

How this classification was reachedexpand

Full frame machine prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.

metaresearch head score (Codex)0.001
metaresearch head score (Gemma)0.001
Version: metacan-v3-hybrid-931329e0061cValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.008
Threshold uncertainty score0.015

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0010.001
Meta-epidemiology (narrow)0.0010.000
Meta-epidemiology (broad)0.0010.000
Bibliometrics0.0000.001
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0010.000
Research integrity0.0000.001
Insufficient payload (model declined to judge)0.0020.002

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.023
GPT teacher head0.285
Teacher spread0.263 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreEmpirical

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

Quick stats

Citations0
Published2014
Admission routes1
Has abstractyes

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