High-resolution structure of the heat-stable form-IAq RuBisCO from the thermophilic purple sulfur bacterium Thermochromatium tepidum

Abstract Ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO) catalyzes the initial carbon fixation reaction in the Calvin-Benson-Bassham cycle. Among the many forms of RuBisCOs, form-I—a protein complex containing 8 large and 8 small subunits—is the most common, representing over 90% of all kn...

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Main Authors: Shenghai Chang, Weiwei Wang, Michael T. Madigan, Long-Jiang Yu, Haichun Gao, Zheng‑Yu Wang‑Otomo, Xing Zhang, Jing-Hua Chen
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-07081-8
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author Shenghai Chang
Weiwei Wang
Michael T. Madigan
Long-Jiang Yu
Haichun Gao
Zheng‑Yu Wang‑Otomo
Xing Zhang
Jing-Hua Chen
author_facet Shenghai Chang
Weiwei Wang
Michael T. Madigan
Long-Jiang Yu
Haichun Gao
Zheng‑Yu Wang‑Otomo
Xing Zhang
Jing-Hua Chen
author_sort Shenghai Chang
collection DOAJ
description Abstract Ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO) catalyzes the initial carbon fixation reaction in the Calvin-Benson-Bassham cycle. Among the many forms of RuBisCOs, form-I—a protein complex containing 8 large and 8 small subunits—is the most common, representing over 90% of all known RuBisCOs. Although many form-I RuBisCO structures have been determined, no structure has been reported for a form-IAq RuBisCO. Here, we detail the structure of the heat-stable form-IAq RuBisCO from the thermophilic and anaerobic purple bacterium Thermochromatium (Tch.) tepidum at 1.55 Å resolution. The overall structure of the Tch. tepidum form-IAq RuBisCO resembles both a form-IAc RuBisCO from a chemolithotrophic sulfur bacterium and a synthetic form-I RuBisCO reconstructed from ancestral sequences. However, the Tch. tepidum enzyme shows significantly greater interactions between adjacent small subunits through their extended N-terminal domains that contain a characteristic six-residue insertion unique to form-IAq RuBisCOs. Structural differences of Tch. tepidum RuBisCO from its mesophilic relative Allochromatium vinosum, and key substitutions on the hydrophilic surface of the small subunits suggests the mechanisms of its enhanced thermostability. Our structure represents the first structure of a form-IAq RuBisCO, providing fresh clues for unraveling the evolutionary history of RuBisCO and new details for how this key enzyme remains active at elevated temperatures.
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spelling doaj-art-791d2ee22ca9401890d11f1c494ca85a2025-08-20T03:38:12ZengNature PortfolioScientific Reports2045-23222025-07-0115111110.1038/s41598-025-07081-8High-resolution structure of the heat-stable form-IAq RuBisCO from the thermophilic purple sulfur bacterium Thermochromatium tepidumShenghai Chang0Weiwei Wang1Michael T. Madigan2Long-Jiang Yu3Haichun Gao4Zheng‑Yu Wang‑Otomo5Xing Zhang6Jing-Hua Chen7Department of Biophysics, Department of Pathology of Sir Run Run Shaw Hospital, Zhejiang University School of MedicineCollege of Life Sciences, Zhejiang UniversitySchool of Biological Sciences, Department of Microbiology, Southern Illinois UniversityPhotosynthesis Research Center, Key Laboratory of Photobiology, Institute of Botany, Chinese Academy of SciencesCollege of Life Sciences, Zhejiang UniversityFaculty of Science, Ibaraki UniversityDepartment of Biophysics, Department of Pathology of Sir Run Run Shaw Hospital, Zhejiang University School of MedicineCollege of Life Sciences, Zhejiang UniversityAbstract Ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO) catalyzes the initial carbon fixation reaction in the Calvin-Benson-Bassham cycle. Among the many forms of RuBisCOs, form-I—a protein complex containing 8 large and 8 small subunits—is the most common, representing over 90% of all known RuBisCOs. Although many form-I RuBisCO structures have been determined, no structure has been reported for a form-IAq RuBisCO. Here, we detail the structure of the heat-stable form-IAq RuBisCO from the thermophilic and anaerobic purple bacterium Thermochromatium (Tch.) tepidum at 1.55 Å resolution. The overall structure of the Tch. tepidum form-IAq RuBisCO resembles both a form-IAc RuBisCO from a chemolithotrophic sulfur bacterium and a synthetic form-I RuBisCO reconstructed from ancestral sequences. However, the Tch. tepidum enzyme shows significantly greater interactions between adjacent small subunits through their extended N-terminal domains that contain a characteristic six-residue insertion unique to form-IAq RuBisCOs. Structural differences of Tch. tepidum RuBisCO from its mesophilic relative Allochromatium vinosum, and key substitutions on the hydrophilic surface of the small subunits suggests the mechanisms of its enhanced thermostability. Our structure represents the first structure of a form-IAq RuBisCO, providing fresh clues for unraveling the evolutionary history of RuBisCO and new details for how this key enzyme remains active at elevated temperatures.https://doi.org/10.1038/s41598-025-07081-8PhotosynthesisRuBisCOThermochromatium tepidumCryo-EM
spellingShingle Shenghai Chang
Weiwei Wang
Michael T. Madigan
Long-Jiang Yu
Haichun Gao
Zheng‑Yu Wang‑Otomo
Xing Zhang
Jing-Hua Chen
High-resolution structure of the heat-stable form-IAq RuBisCO from the thermophilic purple sulfur bacterium Thermochromatium tepidum
Scientific Reports
Photosynthesis
RuBisCO
Thermochromatium tepidum
Cryo-EM
title High-resolution structure of the heat-stable form-IAq RuBisCO from the thermophilic purple sulfur bacterium Thermochromatium tepidum
title_full High-resolution structure of the heat-stable form-IAq RuBisCO from the thermophilic purple sulfur bacterium Thermochromatium tepidum
title_fullStr High-resolution structure of the heat-stable form-IAq RuBisCO from the thermophilic purple sulfur bacterium Thermochromatium tepidum
title_full_unstemmed High-resolution structure of the heat-stable form-IAq RuBisCO from the thermophilic purple sulfur bacterium Thermochromatium tepidum
title_short High-resolution structure of the heat-stable form-IAq RuBisCO from the thermophilic purple sulfur bacterium Thermochromatium tepidum
title_sort high resolution structure of the heat stable form iaq rubisco from the thermophilic purple sulfur bacterium thermochromatium tepidum
topic Photosynthesis
RuBisCO
Thermochromatium tepidum
Cryo-EM
url https://doi.org/10.1038/s41598-025-07081-8
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