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Record W4390342007 · doi:10.1002/pro.4882

<scp><i>Helicobacter pylori</i> FlgN</scp> binds its substrate <scp>FlgK</scp> and the flagellum <scp>ATPase FliI</scp> in a similar manner observed for the <scp>FliT</scp> chaperone

2023· article· en· W4390342007 on OpenAlexafffund
Poonam Dhindwal, Michal T. Boniecki, S A Moore

Bibliographic record

VenueProtein Science · 2023
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicEscherichia coli research studies
Canadian institutionsUniversity of Saskatchewan
FundersNatural Sciences and Engineering Research Council of CanadaCanadian Institutes of Health Research
KeywordsFlagellumChaperone (clinical)BiologyATPaseSecretionMicroscale thermophoresisCell biologyBiophysicsBiochemistryGeneEnzyme

Abstract

fetched live from OpenAlex

Abstract In bacterial flagellum biogenesis, secretion of the hook–filament junction proteins FlgK and FlgL and completion of the flagellum requires the FlgN chaperone. Similarly, the related FliT chaperone is necessary for the secretion of the filament cap protein FliD and binds the flagellar export gate protein FlhA and the flagellum ATPase FliI. FlgN and FliT require FliJ for effective substrate secretion. In Helicobacter pylori , neither FlgN, FliT, nor FliJ have been annotated. We demonstrate that the genome location of HP1120 is identical to that of flgN in other flagellated bacteria and that HP1120 is the homolog of Campylobacter jejuni FlgN. A modeled HP1120 structure contains three α‐helices and resembles the FliT chaperone, sharing a similar substrate‐binding pocket. Using pulldowns and thermophoresis, we show that both HP1120 and a HP1120Δ 126–144 deletion mutant bind to FlgK with nanomolar affinity, but not to the filament cap protein FliD, confirming that HP1120 is FlgN. Based on size‐exclusion chromatography and multi‐angle light scattering, H. pylori FlgN binds to FlgK with 1:1 stoichiometry. Overall structural similarities between FlgN and FliT suggest that substrate recognition on FlgN primarily involves an antiparallel coiled‐coil interface between the third helix of FlgN and the C‐terminal helix of the substrate. A FlgNΔ 126–144 N100A, Y103A, S111I triple mutant targeting this interface significantly impairs the binding of FlgK. Finally, we demonstrate that FlgNΔ 126–144 , like FliT, binds with sub‐micromolar affinity to the flagellum ATPase FliI or its N‐terminal domain. Hence FlgN and FliT likely couple delivery of low‐abundance export substrates to the flagellum ATPase FliI.

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)0.007
metaresearch head score (Gemma)0.017
Version: codex-gemma-dda1882f352aValidation status: machine_predicted_unvalidated
Candidate categoriesMetaresearch, Meta-epidemiology (narrow), Science and technology studies
Consensus categoriesScience and technology studies
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 score0.277
Threshold uncertainty score1.000

Codex and Gemma teacher scores by category

CategoryCodexGemma
Metaresearch0.0070.017
Meta-epidemiology (narrow)0.0010.001
Meta-epidemiology (broad)0.0010.000
Bibliometrics0.0000.003
Science and technology studies0.0020.003
Scholarly communication0.0010.000
Open science0.0030.002
Research integrity0.0000.001
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 head0.038
GPT teacher head0.279
Teacher spread0.241 · 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; both teacher heads agree on what is shown here.

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

Citations1
Published2023
Admission routes2
Has abstractyes

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