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Record W2078253671 · doi:10.1177/1087057111415525

Discovery of Novel P2Y14 Agonist and Antagonist Using Conventional and Nonconventional Methods

2011· article· en· W2078253671 on OpenAlex

Why this work is in the frame

A frame that forgets how it found something cannot be audited. These are the routes that admitted this work.

affAt least one author lists a Canadian institution in the pinned OpenAlex snapshot.

Bibliographic record

VenueSLAS DISCOVERY · 2011
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicAdenosine and Purinergic Signaling
Canadian institutionsMerck Canada Inc. (Canada)
Fundersnot available
KeywordsAgonistAntagonistPharmacologyChemistryMedicineInternal medicineReceptor

Abstract

fetched live from OpenAlex

P2Y14 is a member of the pyrimidinergic GPCR family. UDP-Glc has been previously shown to activate human P2Y14, whereas UDP was unable to activate the receptor. In this study, the authors used conventional and nonconventional methods to further characterize P2Y14 and its ligands. Conventional calcium mobilization and nonconventional cellular impedance functional assays revealed that UMP and UDP selectively activated HEK cells coexpressing P2Y14 and Gαqi5. In the impedance assays, the presence of exogenous Gαqi5 resulted in agonist-induced Gq signaling, whereas in the absence of exogenous Gαqi5, the signal was indicative of Gi. The authors established the first P2Y14 membrane filtration binding assay using a novel optimized expression vector and [3H]UDP as radioligand. UDP-Glc, UMP, and UDP dose dependently inhibited [3H]UDP binding in the binding assay, and saturation analysis revealed that UDP bound P2Y14 with a KD = 10 nM and a Bmax = 110 pmol/mg. The authors screened a phosphonate library and identified compound A, which inhibited UDP-Glc–mediated calcium signaling in the fluorometric imaging plate reader assay (IC50 = 2.3 µM) and competed for [3H]UDP binding in the novel binding assay with a Ki = 1280 nM. P2Y14 is a member of the pyrimidinergic GPCR family. UDP-Glc has been previously shown to activate human P2Y14, whereas UDP was unable to activate the receptor. In this study, the authors used conventional and nonconventional methods to further characterize P2Y14 and its ligands. Conventional calcium mobilization and nonconventional cellular impedance functional assays revealed that UMP and UDP selectively activated HEK cells coexpressing P2Y14 and Gαqi5. In the impedance assays, the presence of exogenous Gαqi5 resulted in agonist-induced Gq signaling, whereas in the absence of exogenous Gαqi5, the signal was indicative of Gi. The authors established the first P2Y14 membrane filtration binding assay using a novel optimized expression vector and [3H]UDP as radioligand. UDP-Glc, UMP, and UDP dose dependently inhibited [3H]UDP binding in the binding assay, and saturation analysis revealed that UDP bound P2Y14 with a KD = 10 nM and a Bmax = 110 pmol/mg. The authors screened a phosphonate library and identified compound A, which inhibited UDP-Glc–mediated calcium signaling in the fluorometric imaging plate reader assay (IC50 = 2.3 µM) and competed for [3H]UDP binding in the novel binding assay with a Ki = 1280 nM.

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.

Full frame distilled prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.

metaresearch head score (Codex)0.000
metaresearch head score (Gemma)0.000
Version: codex-gemma-dda1882f352aValidation 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 score0.113
Threshold uncertainty score0.567

Codex and Gemma teacher scores by category

CategoryCodexGemma
Metaresearch0.0000.000
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.000
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0000.000
Research integrity0.0000.000
Insufficient payload (model declined to judge)0.0000.000

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 head0.048
GPT teacher head0.320
Teacher spread0.273 · 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