A disulfide redox switch mechanism regulates glycoside hydrolase function
Disulfide bonds are a key post-translational modification involved in protein folding, structural stability, and functional regulation. Here, we demonstrate that a glycoside hydrolase from the GH2 family undergoes reversible redox regulation through an intramolecular disulfide bond. The enzyme is inactive in its oxidized state and becomes active when reduced through a fully reversible process. Under oxidative conditions, multiple crystallographic and cryo-EM structures revealed a pronounced structural disorder in the active site, most prominent in the regulatory and catalytic loops, which disrupts the substrate binding site and, remarkably, the configuration of the acidic catalytic residues. Conversely, a high-resolution cryo-EM structure of the active (reduced) state unveiled a well-ordered active site with catalytic residues properly positioned for a classical Koshland retaining mechanism. This reversible order-disorder process based on a disulfide switch provides a mechanism for redox-dependent control of glycoside hydrolase activity, with potential implications for carbohydrate metabolism, microbial adaptation and biotechnological applications.
Authors
- Yoshihisa Yoshimi (ORCID: https://orcid.org/0000-0002-6734-6677)
- Luciano G. Dolce (ORCID: https://orcid.org/0000-0003-0871-297X)
- Renan Y. Miyamoto (ORCID: https://orcid.org/0000-0003-2990-3337)
- Rodrigo Silva Araujo Streit (ORCID: https://orcid.org/0009-0001-7518-5201)
- Gabriela Félix Persinoti (ORCID: https://orcid.org/0000-0002-0975-7283)
- Si Li (ORCID: https://orcid.org/0000-0002-5431-1301)
- Paul Dupree (ORCID: https://orcid.org/0000-0001-9270-6286)
- M.T. Murakami (ORCID: https://orcid.org/0000-0002-0405-8010)
- João Paulo M. Spadeto (ORCID: https://orcid.org/0000-0002-4193-8797)
- Marcele Pandeló Martins (ORCID: https://orcid.org/0000-0002-5015-7116)
- Gustavo Henrique Martins (ORCID: https://orcid.org/0000-0003-3749-1520)
- António Carlos Borges
- Fabiane Stoffel
- Mariana Abrahão Bueno Morais
- Rafael Henrique Galinari
- Felipe Jun Fuzita
- Camila Ramos dos Santos
Institutions
- Centre National de la Recherche Scientifique (FR)
- Inserm (FR)
- Universidade Estadual de Campinas (UNICAMP) (BR)
- University of Cambridge (GB)
- Cambridge School (PT)
- Bridge University (SS)
- Brazilian Center for Research in Energy and Materials (BR)
- Institut pour l'avancée des biosciences (FR)
- Université Grenoble Alpes (FR)
Publication Details
- Journal
- Nature Communications
- Published
- 2026-01-05
- DOI
- https://doi.org/10.1038/s41467-025-67225-2
- Citations
- 1
- Primary Topic
- Redox biology and oxidative stress
- Type
- article
- Field-Weighted Citation Impact
- 22.24
Funders
- Fundação de Amparo à Pesquisa do Estado de São Paulo
- Ministério da Ciência, Tecnologia e Inovação
- Conselho Nacional de Desenvolvimento Científico e Tecnológico
- Laboratório Nacional de Luz Síncrotron
- Laboratório Nacional de Biociências
- Laboratório Nacional de Nanotecnologia
- Laboratório Nacional de Biorrenováveis
- SLAC National Accelerator Laboratory