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Record W4416714296 · doi:10.1038/s42004-025-01763-0

Challenging AlphaFold in predicting proteins with large-scale allosteric transitions

2025· article· en· W4416714296 on OpenAlexafffund
Brooks H. Perkins-Jechow, Juan Pablo Iglesias Ahualli, Huyen Thuc Nhu, Alireza Omidi, C. Xue Li, Jorge A. Holguin-Cruz, Dokyun Na, Nawar Malhis, Jennifer M. Bui, Jörg Gsponer

Bibliographic record

VenueCommunications Chemistry · 2025
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicProtein Structure and Dynamics
Canadian institutionsMichael Smith Health Research BCUniversity of British Columbia HospitalCanada's Michael Smith Genome Sciences CentreUniversity of British Columbia
FundersNatural Sciences and Engineering Research Council of CanadaAlliance de recherche numérique du CanadaCanadian Network for Research and Innovation in Machining Technology, Natural Sciences and Engineering Research Council of Canada
KeywordsAllosteric regulationSequence (biology)Class (philosophy)Function (biology)Protein structureConformational ensembles

Abstract

fetched live from OpenAlex

Many proteins function by toggling between distinct conformations, yet most structure predictors have been trained on data that do not capture this conformational diversity. Here, we benchmarked AlphaFold2, AlphaFold3, and recent variants on autoinhibited proteins, a class of allosterically regulated, often multi-domain proteins that exist in equilibrium between active and autoinhibited states. Our analyses show that AlphaFold2 fails to reproduce the experimental structures of many autoinhibited proteins, which is reflected in reduced confidence scores. This contrasts sharply with its high-accuracy, high-confidence predictions of non-autoinhibited multi-domain proteins. When tested for its ability to capture conformational diversity, we found that AlphaFold2 performs better when combined with uniform subsampling of sequence alignments rather than local subsampling. BioEmu and AlphaFold3 improve upon these results, yet still struggle to accurately reproduce details of experimental structures. Together, our study underscores the persistent challenges of predicting protein structures shaped by complex energy landscapes. Although many proteins function by toggling between distinct conformations, most structure predictors remain limited to a single static fold. Here, the authors test the performance of AlphaFold2, AlphaFold3, and recent variants on a dataset of autoinhibited proteins exhibiting at least two functionally distinct conformations, and show that AlphaFold2 fails to reproduce the experimental structures of many autoinhibited proteins, but that it can capture conformational diversity when using uniform multiple sequence alignment subsampling.

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 score0.122
Threshold uncertainty score0.514

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 science0.0010.000
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 head0.006
GPT teacher head0.245
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

Citations8
Published2025
Admission routes2
Has abstractyes

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