KinForm: kinetics-informed feature optimised representation models for enzyme kcat and KM prediction

Kinetic parameters such as the turnover number (k<sub>cat</sub>) and Michaelis constant (K<sub>M</sub>) are essential for modelling enzymatic activity but experimental data remains limited in scale and diversity. Previous methods for predicting enzyme kinetics typically use mean-pooled residue embeddings from a single protein language model to represent the protein. We present KinForm, a machine learning framework designed to improve predictive accuracy and generalisation for kinetic parameters by optimising protein feature representations. KinForm combines several residue-level embeddings (Evolutionary Scale Modelling Cambrian, Evolutionary Scale Modelling 2, and ProtT5-XL-UniRef50), taken from empirically selected intermediate transformer layers, and applies weighted pooling based on per-residue binding-site probability. To counter the resulting high dimensionality, we apply dimensionality reduction using principal-component analysis (PCA) on concatenated protein features, and rebalance the training data via a similarity-based oversampling strategy. KinForm outperforms baseline methods on two benchmark datasets. Improvements are most pronounced in low sequence similarity bins. We observe improvements from binding-site probability pooling, intermediate-layer selection, PCA, and oversampling of low-identity proteins. We also find that removing sequence overlap between folds provides a more realistic evaluation of generalisation and should be the standard over random splitting when benchmarking kinetic prediction models.

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Publication Details

Journal
npj Systems Biology and Applications
Published
2026-03-28
DOI
https://doi.org/10.1038/s41540-026-00692-5
Primary Topic
Protein Structure and Dynamics
Type
article
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article

KinForm: kinetics-informed feature optimised representation models for enzyme kcat and KM prediction

Ronan M. T. Fleming, Saleh Alwer
npj Systems Biology and Applications
Protein Structure and Dynamics
article

KinForm: kinetics-informed feature optimised representation models for enzyme kcat and KM prediction

Ronan M. T. Fleming, Saleh Alwer
article en

Abstract

Kinetic parameters such as the turnover number (k<sub>cat</sub>) and Michaelis constant (K<sub>M</sub>) are essential for modelling enzymatic activity but experimental data remains limited in scale and diversity. Previous methods for predicting enzyme kinetics typically use mean-pooled residue embeddings from a single protein language model to represent the protein. We present KinForm, a machine learning framework designed to improve predictive accuracy and generalisation for kinetic parameters by optimising protein feature representations. KinForm combines several residue-level embeddings (Evolutionary Scale Modelling Cambrian, Evolutionary Scale Modelling 2, and ProtT5-XL-UniRef50), taken from empirically selected intermediate transformer layers, and applies weighted pooling based on per-residue binding-site probability. To counter the resulting high dimensionality, we apply dimensionality reduction using principal-component analysis (PCA) on concatenated protein features, and rebalance the training data via a similarity-based oversampling strategy. KinForm outperforms baseline methods on two benchmark datasets. Improvements are most pronounced in low sequence similarity bins. We observe improvements from binding-site probability pooling, intermediate-layer selection, PCA, and oversampling of low-identity proteins. We also find that removing sequence overlap between folds provides a more realistic evaluation of generalisation and should be the standard over random splitting when benchmarking kinetic prediction models.

npj Systems Biology and Applications
Ollscoil na Gaillimhe – University of Galway (IE), COPE Galway (IE), Trials Methodology Research Network (IE)
Openalex Percentile: Top 5%
Protein Structure and Dynamics
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