دورية أكاديمية
The Knotted Protein UCH-L1 Exhibits Partially Unfolded Forms under Native Conditions that Share Common Structural Features with Its Kinetic Folding Intermediates.
| العنوان: | The Knotted Protein UCH-L1 Exhibits Partially Unfolded Forms under Native Conditions that Share Common Structural Features with Its Kinetic Folding Intermediates. |
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| المؤلفون: | Lou, Shih-Chi, Wetzel, Svava, Zhang, Hongyu, Crone, Elizabeth W, Lee, Yun-Tzai, Jackson, Sophie E, Hsu, Shang-Te Danny |
| بيانات النشر: | Elsevier BV //dx.doi.org/10.1016/j.jmb.2016.04.002 J Mol Biol |
| سنة النشر: | 2016 |
| المجموعة: | Apollo - University of Cambridge Repository |
| مصطلحات موضوعية: | folding intermediate, folding pathway, knotted protein, partially unfolded form, ubiquitin C-terminal hydrolase, Deuterium, Deuterium Exchange Measurement, Humans, Hydrogen, Hydrogen-Ion Concentration, Kinetics, Magnetic Resonance Spectroscopy, Protein Conformation, alpha-Helical, beta-Strand, Protein Denaturation, Protein Folding, Ubiquitin Thiolesterase |
| الوصف: | The human ubiquitin C-terminal hydrolase, UCH-L1, is an abundant neuronal deubiquitinase that is associated with Parkinson's disease. It contains a complex Gordian knot topology formed by the polypeptide chain alone. Using a combination of fluorescence-based kinetic measurements, we show that UCH-L1 has two distinct kinetic folding intermediates that are transiently populated on parallel pathways between the denatured and native states. NMR hydrogen-deuterium exchange (HDX) experiments indicate the presence of partially unfolded forms (PUFs) of UCH-L1 under native conditions. HDX measurements as a function of urea concentration were used to establish the structure of the PUFs and pulse-labelled HDX NMR was used to show that the PUFs and the folding intermediates are likely the same species. In both cases, a similar stable core encompassing most of the central β-sheet is highly structured and α-helix 3, which is partially formed, packs against it. In contrast to the stable β-sheet core, the peripheral α-helices display significant local fluctuations leading to rapid exchange. The results also suggest that the main difference between the two kinetic intermediates is structure and packing of α-helices 3 and 7 and the degree of structure in β-strand 5. Together, the fluorescence and NMR results establish that UCH-L1 neither folds through a continuum of pathways nor by a single discrete pathway. Its folding is complex, the β-sheet core forms early and is present in both intermediate states, and the rate-limiting step which is likely to involve the threading of the chain to form the 52-knot occurs late on the folding pathway. ; This work was supported by the National Science Council (99-2911-I-007-034 and 104-2113-M-001-016), National Tsing Hua University and Academia Sinica, Taiwan. S.-T.D.H. was supported by a Career Development Award (CDA- 00025/2010-C) from the International Human Frontier Science Program. The NMR spectra were obtained at the NMR facility of the Department of Chemistry, University of Cambridge and ... |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | application/pdf |
| اللغة: | English |
| العلاقة: | https://www.repository.cam.ac.uk/handle/1810/255728Test |
| الإتاحة: | https://www.repository.cam.ac.uk/handle/1810/255728Test |
| حقوق: | Attribution-NonCommercial-NoDerivatives 4.0 International ; http://creativecommons.org/licenses/by-nc-nd/4.0Test/ |
| رقم الانضمام: | edsbas.D6F9E9BB |
| قاعدة البيانات: | BASE |
| الوصف غير متاح. |