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Thermostable chaperonin from Clostridium thermocellum
S J Cross1, A Ciruela, K Poomputsa
1Research School of Biosciences, University of Kent at Canterbury, U.K.
The Biochemical Journal
|June 1, 1996
Summary
Researchers purified chaperonins Cpn60 and Cpn10 from Clostridium thermocellum. The Cpn60 homologue is a thermostable ATPase, showing structural similarities to E. coli GroEL.
Area of Science:
- Molecular Biology
- Biochemistry
- Microbiology
Background:
- Chaperonins are essential molecular machines involved in protein folding.
- Clostridium thermocellum is a thermophilic anaerobe with industrial relevance in biomass degradation.
Purpose of the Study:
- To purify and characterize chaperonins Cpn60 and Cpn10 from Clostridium thermocellum.
- To investigate the ATPase activity and quaternary structure of C. thermocellum Cpn60.
Main Methods:
- ATP-affinity chromatography for Cpn60 purification.
- Gel-filtration, ion-exchange, and hydrophobic interaction chromatography for Cpn10 purification.
- N-terminal sequencing, antigenic cross-reactivity, ATPase assays, gel-filtration, and electron microscopy for characterization.
Main Results:
- Cpn60 and Cpn10 homologues were successfully purified from C. thermocellum.
- C. thermocellum Cpn60 exhibits weak, thermostable ATPase activity with optimal function between 60-70°C.
- Electron microscopy revealed Cpn60 exists mainly as a heptameric complex with 7-fold symmetry, and some tetradecamers, structurally resembling E. coli GroEL.
Conclusions:
- The study successfully characterized Cpn60 and Cpn10 from C. thermocellum, providing insights into their biochemical and structural properties.
- The findings suggest functional and structural conservation of chaperonins across different bacterial species, including thermophiles.