ORIGINAL RESEARCH article

Front. Chem.

Sec. Theoretical and Computational Chemistry

ESM3-Driven Hybrid Design of a Hemoglobin Variant with Enhanced Functionality and Structural Stability using Protein Language Model

  • 1. ITMO University, Saint Petersburg, Russia

  • 2. Julius-Maximilians-Universitat Wurzburg, Würzburg, Germany

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Abstract

This study presents a hybrid computational pipeline integrating rational protein design with the ESM3 multimodal generative model to validate a known high-oxygen-affinity hemoglobin (Hb) variant (Hb Fort de France, α45[CD3]His→Arg). Using structural homology to the Hb heme-binding site, we generated an esmHb model with the targeted His45Arg substitution via ESM3. Analyses through integrated computational methods, including Monte Carlo and molecular dynamics simulations, molecular docking, FoldX, MM-GBSA/PBSA, and DFT revealed enhanced structural stability, stronger heme binding (MM-GBSA ΔG: -29.28 vs. -21.75 kcal/mol; MM-PBSA: -64.14 vs. -51.00 kcal/mol), improved subunit interactions, and more favorable oxidation thermodynamics (ΔE improvement: 23.2 kcal/mol) compared to wild-type Hb. These results suggest the utility of the ESM3 model for precise benchmarking and function-preserving engineering of therapeutically relevant proteins like optimized Hb for oxygen therapeutics.

Summary

Keywords

Computational Protein Design, DFT calculations, Hemoglobin, molecular dynamics, Monte Carlo simulation, Myoglobin, oxygen affinity, protein language model

Received

02 June 2026

Accepted

31 July 2026

Copyright

© 2026 Shityakov, Novikov, Skorb and Dandekar. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

*Correspondence: Sergey Shityakov

Disclaimer

All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.

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