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Record W2127133496 · doi:10.1074/jbc.a114.607359

HD1, a thrombin-directed aptamer, binds exosite 1 on prothrombin with high affinity and inhibits its activation by prothrombinase.

2015· article· en· W2127133496 on OpenAlexaff
Colin A. Kretz, Alan R. Stafford, James C. Fredenburgh, Jeffrey I. Weitz

Bibliographic record

VenueJournal of Biological Chemistry · 2015
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicAdvanced biosensing and bioanalysis techniques
Canadian institutionsMcMaster University Medical Centre
Fundersnot available
KeywordsAptamerThrombinChemistryOligonucleotideSurface plasmon resonanceBiophysicsStreptavidinMolecular biologyBiochemistryStereochemistryDNABiologyNanotechnologyBiotin

Abstract

fetched live from OpenAlex

VOLUME 281 (2006) PAGES 37477–37485 The sequences reported for the HD22 and HD23 aptamers were not correct. 1) The aptamer that we called HD23, an inactive variant of HD22 that does not bind thrombin previously named ODN 4a (Dougan, H., Weitz, J. I., Stafford, A. R., Gillespie, K. D., Klement, P., Hobbs, J. B., and Lyster, D. M. (2003) Evaluation of DNA aptamers directed to thrombin as potential thrombus imaging agents. Nucl. Med. Biol. 30, 61–72), was reported with a sequence that was missing three nucleotides. We are now calling this aptamer SV23, as shown below. However, HD23 was used in this study.AGTCCGTAATAAAGCAGGTTAAAATGACT(HD23) AGTCCG - - -TAAAGCAGGTTAAAATGACT(SV23) 2) Instead of using the aptamer HD22 that was previously named (60–18[29]) and was reported to bind thrombin (Tasset, D. M., Kubik, M. F., and Steiner, W. (1997) Oligonucleotide inhibitors of human thrombin that bind distinct epitopes. J. Mol. Biol. 272, 688–698), we used an aptamer in which the TGG triplet at position 7–9 was duplicated. We are now calling this aptamer EV22, as shown below.AGTCCGTGG - - -TAGGGCAGGTTGGGGTGACT(HD22) AGTCCGTGGTGGTAGGGCAGGTTGGGGTGACT(EV22) To determine how this error may have affected our results, we used surface plasmon resonance to compare the thrombin binding affinity of EV22 with that of HD22. Biotin-labeled aptamers were adsorbed onto separate streptavidin-modified flow cells and active thrombin was then injected. Analyses of on- and off-rates yielded Kd values of 39 nm and 0.3 nm for EV22 and HD22, respectively. Similar values were obtained when Phe-Pro-Arg-chloromethyl ketone-inhibited thrombin was injected in place of active thrombin. The Kd value of 0.3 nm for thrombin binding to HD22 is consistent with the Kd value of 0.5 nm reported previously for 60–18[29] (Tasset et al.). Thus, EV22 exhibited 130-fold weaker thrombin binding affinity than HD22. γ-Thrombin, which lacks exosite 1, bound EV22, whereas R93E thrombin, a variant with an impaired exosite 2, did not bind EV22, so EV22 appears to exhibit the same exosite 2 specificity as HD22. Because EV22 differs from HD22 in affinity but not specificity, we conclude that these errors do not affect the interpretation of the results or the conclusions of this work.

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 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 score0.002
Threshold uncertainty score0.671

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 head0.023
GPT teacher head0.261
Teacher spread0.239 · 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 teacher head, 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

Citations53
Published2015
Admission routes1
Has abstractyes

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