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Folding of CFTR is predominantly cotranslational
Bertrand Kleizen1, Thijs van Vlijmen, Hugo R de Jonge
1Cellular Protein Chemistry, Department of Chemistry, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
Molecular Cell
|October 26, 2005
Summary
Newly synthesized cystic fibrosis transmembrane conductance regulator (CFTR) protein folds during its creation in the endoplasmic reticulum. Individual protein domains form structures independently, suggesting a step-by-step folding process.
Area of Science:
- Molecular biology
- Cell biology
- Protein folding
Background:
- The folding mechanisms of newly synthesized, multispanning membrane proteins in the endoplasmic reticulum (ER) remain largely uncharacterized.
- Understanding protein folding is crucial for cellular function and disease mechanisms.
Purpose of the Study:
- To investigate the early folding events of the cystic fibrosis transmembrane conductance regulator (CFTR) during its synthesis.
- To determine if CFTR folding occurs cotranslationally or post-translationally.
Main Methods:
- In vitro translation of CFTR in semipermeabilized cells to study nascent chain elongation.
- Protease susceptibility assays to assess protein folding.
- Analysis of C-terminally truncated CFTR constructs.
Main Results:
- CFTR folding was found to occur predominantly during synthesis, as indicated by protease susceptibility.
- Individual domains of CFTR formed stable, well-defined structures.
- These structures formed independently of other C-terminal parts of the protein.
Conclusions:
- The multidomain protein CFTR undergoes primarily cotranslational folding.
- Folding occurs in a stepwise manner, with individual domains adopting structures during synthesis.