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Record W2058084745 · doi:10.1016/j.jalz.2012.05.1257

P3‐039: Aβ binding properties of a synthetic peptide corresponding to an amino acid sequence in human pericentriolar material 1 (PCM‐1) protein

2012· article· en· W2058084745 on OpenAlexaff
Balu Chakravarthy, Trevor Atkinson, Leslie Brown, Michel Ménard, Shingo Ito, James Whitfield

Bibliographic record

VenueAlzheimer s & Dementia · 2012
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
Topic14-3-3 protein interactions
Canadian institutionsNational Research Council Canada
Fundersnot available
KeywordsMolecular biologyPeptide sequencePeptideBiochemistryAmino acidWestern blotBinding proteinPolyclonal antibodiesBiologyChemistryAntibody

Abstract

fetched live from OpenAlex

We have isolated a ∼ 20kDa polypeptide (termed PK-4) from a T-7 Phage Display system expressing human brain cDNA library (Novagen) using polyclonal antibodies against a protein kinase C (PKC) inhibitor as bait. In addition to having PKC inhibitory activity, we serendipitously discovered that this polypeptide also bound Aß 1–42 with high affinity. The amino acid sequence of this polypeptide has 89 percent homology to an amino-acid sequence in human pericentriolar material-1 (PCM-1) protein. We have mapped the Aß-binding motif on this polypeptide, and have synthesized short (∼4 kDA; 40 amino acids) amyloid-binding peptides (ABPs) corresponding to this region. We have determined the Aß 1–42 binding properties of one of these ABPs, ABP-p4–5, using ELISA, dot blot and poly-acrylamide gel electrophoresis, and immunohistochemistry with brain sections. Cell toxicity assays were done with SH-SY5Y human neuroblastoma cells and Hoechst stain. We have found that ABP-p4–5 binds Aß 1–42 in the nM range. Thioflavin-binding assay and Western blot analysis indicate that ABP-p4–5 inhibits Aß 1–42 aggregation. Interestingly, ELISA studies indicated that ABP-p4–5 bound aggregated Aß1–42 preparations (37°C, 24 hrs) better than non-aggregated preparations, suggesting that it may have higher affinity towards Aß oligomers. In cell toxicity assays using human SH-SY5Y neuroblastoma cells, ABP-p4–5 inhibited Aß1–42-induced apoptotic cell death. Furthermore, we have found using fluorophore labeled p4–5 (FITC-p4–5) that it binds amyloid deposits in brain sections from Alzheimer's disease (AD)-transgenic mice and human AD patients in vitro and also when microinjected into AD-mouse brain. These results indicate that the ABP-p4–5 peptide can potentially be used therapeutically to reduce Aß burden and also to detect Aß deposits in the brain. Most importantly, since PCM-1 and the Aß 1–42-binding ABP-p4–5 have a high-sequence homology it would be interesting to determine whether PCM-1 has a role in the development of AD. Thus, for example, PCM-1 is known to be required for ciliogenesis which in turn is known to be required for neurogenesis and memory formation. So the question remains, would the accumulating oligomeric Aß 1–42 bind PCM-1 and prevent it from promoting ciliogenesis and supporting neurogenesis in an AD-developing brain?

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 score0.003
Threshold uncertainty score0.909

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 head0.039
GPT teacher head0.290
Teacher spread0.251 · 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

Citations0
Published2012
Admission routes1
Has abstractyes

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