Thermostable proteins are crucial in numerous biomedical and biotechnological applications. However, naturally occurring proteins have evolved to function in mild conditions, and laboratory experiments aiming at improving protein stability have proven laborious and expensive. Computational methods overcome this issue by providing a cheap and scalable alternative. Despite significant progress, their reliability is still hindered by the availability of high-quality data. FireProtDB 2.0 (http://loschmidt.chemi.muni.cz/fireprotdb) is a large-scale database aggregating stability data from multiple sources. The second version builds upon its predecessor, retaining its original functionality while introducing a new approach to data storage and maintenance. The new scheme enables the introduction of both absolute and relative data types connected with measurements of wild-types, mutants, protein domains, and de novo designed proteins. Furthermore, while the original database was limited to single-point mutations, more complex data such as insertions, deletions, and multiple-point mutations are now available. As a result, the inclusion of large-scale mutagenesis has increased the size of the database from 16 000 to almost 5 500 000 experiments. Moreover, the updated abstract scheme is fully expandable with any new measurements and annotations without the need for any restructuring. Finally, the tracking of history together with fixed identifiers is in accordance with the FAIR principles.

Department of Information Systems Faculty of Information Technology Brno University of Technology 612 66 Brno Czech Republic

Instituto de Tecnologia Química e Biológica António Xavier Universidade Nova de Lisboa 2780 157 Oeiras Portugal

International Clinical Research Centre St Anne's University Hospital Brno 602 00 Brno Czech Republic

Jerzy Haber Institute of Catalysis and Surface Chemistry Polish Academy of Sciences PL 30239 Krakow Poland

Loschmidt Laboratories Department of Experimental Biology and RECETOX Masaryk University 625 00 Brno Czech Republic

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