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Exploring the Structural Stability and Assembly Mechanism of Hydrophobin Proteins

2020· article· en· W3016446471 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.
aboutThe title or abstract carries a Canadian signal from the geographic lexicon.

Bibliographic record

VenueThe FASEB Journal · 2020
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicHemoglobin structure and function
Canadian institutionsDalhousie University
Fundersnot available
KeywordsHydrophobinSize-exclusion chromatographySchizophyllum communeChemistryGlobular proteinThioflavinBiophysicsFungal proteinBiochemistryChromatographyBiologyMutantEnzyme

Abstract

fetched live from OpenAlex

Hydrophobins are small, globular proteins with amphiphilic character that are produced and secreted by filamentous fungi. At hydrophobic‐hydrophilic interfaces they self‐assemble into durable amyloid‐containing structures, called rodlets, which create protective, water repellent coatings for fungal spores. Current models of hydrophobin self‐assembly predict that hydrophobin monomers undergo a conformational change at a hydrophobic‐hydrophilic interface and integrate into a growing rodlet, however the mechanistic details of rodlet assembly are unknown. To investigate the assembly mechanism of hydrophobins, we carried out stability studies with SC16, a hydrophobin isolated from Schizophyllum commune. SC16 was recombinantly expressed using E. coli and purified by immobilized Ni 2+ affinity chromatography. NMR spectroscopy was used to determine that the structure of SC16 was minimally perturbed by denaturing (8 M urea) or reducing (2 mM DTT) conditions. Gel filtration chromatography indicated that in solution SC16 exists as a tetramer, suggesting that rodlet assembly does not initiate from hydrophobin monomers. Mutant forms of SC16 are being employed to determine the sequences and conformational changes required for rodlet assembly, with gel filtration chromatography used to determine the multimeric state, thioflavin T assays to quantify amyloid formation, and electron microscopy to visualize rodlet formation. X‐ray crystallography is being used to further characterize the tetrameric structure of SC16. Determining which hydrophobin sequences are responsible for self‐assembly will allow the rational modification of hydrophobins to add new functionalities or influence their self‐assembly. Support or Funding Information NSERC, Dalhousie University Faculty of Medicine, Nova Scotia Research Foundation, BioActives CREATE

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 score0.002
Threshold uncertainty score0.167

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 head0.043
GPT teacher head0.231
Teacher spread0.187 · 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