The human mitochondrial Hsp60 in the APO conformation forms a stable tetradecameric complex
| Autor: | Adrian S. Enriquez, Ricardo A. Bernal, Sudheer K. Molugu, Jay M. Bhatt, Humberto M. Rojo, Zacariah L. Hildenbrand |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
macromolecular substances Biology Human mitochondrial chaperonin law.invention Chaperonin 03 medical and health sciences Adenosine Triphosphate Microscopy Electron Transmission law Report transmission electron microscopy Chaperonin 10 Escherichia coli Humans Nucleotide Protein Structure Quaternary Molecular Biology chemistry.chemical_classification 030102 biochemistry & molecular biology Chaperonin 60 Cell Biology Negative stain Dynamic Light Scattering Recombinant Proteins Mitochondria Folding (chemistry) enzymes and coenzymes (carbohydrates) 030104 developmental biology negative stain chemistry Biochemistry Recombinant DNA Biophysics Protein folding HSP60 Developmental Biology Macromolecule |
| Zdroj: | Cell Cycle |
| ISSN: | 1551-4005 1538-4101 |
| DOI: | 10.1080/15384101.2017.1321180 |
| Popis: | The human mitochondrial chaperonin is a macromolecular machine that catalyzes the proper folding of mitochondrial proteins and is of vital importance to all cells. This chaperonin is composed of 2 distinct proteins, Hsp60 and Hsp10, that assemble into large oligomeric complexes that mediate the folding of non-native polypeptides in an ATP dependent manner. Here, we report the bacterial expression and purification of fully assembled human Hsp60 and Hsp10 recombinant proteins and that Hsp60 forms a stable tetradecameric double-ring conformation in the absence of co-chaperonin and nucleotide. Evidence of the stable double-ring conformation is illustrated by the 15 Å resolution electron microscopy reconstruction presented here. Furthermore, our biochemical analyses reveal that the presence of a non-native substrate initiates ATP-hydrolysis within the Hsp60/10 chaperonin to commence protein folding. Collectively, these data provide insight into the architecture of the intermediates used by the human mitochondrial chaperonin along its protein folding pathway and lay a foundation for subsequent high resolution structural investigations into the conformational changes of the mitochondrial chaperonin. |
| Databáze: | OpenAIRE |
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