التفاصيل البيبلوغرافية
| العنوان: |
Conformational and Structural Characterization of Knotted Proteins. |
| المؤلفون: |
Jeanne Dit Fouque K; Department of Chemistry and Biochemistry and Biomolecular Sciences Institute, Florida International University, Miami, Florida, 33199, United States., Molano-Arevalo JC; Department of Chemistry and Biochemistry and Biomolecular Sciences Institute, Florida International University, Miami, Florida, 33199, United States., Leng F; Department of Chemistry and Biochemistry and Biomolecular Sciences Institute, Florida International University, Miami, Florida, 33199, United States., Fernandez-Lima F; Department of Chemistry and Biochemistry and Biomolecular Sciences Institute, Florida International University, Miami, Florida, 33199, United States. |
| المصدر: |
Biochemistry [Biochemistry] 2024 Sep 17; Vol. 63 (18), pp. 2293-2299. Date of Electronic Publication: 2024 Aug 27. |
| نوع المنشور: |
Journal Article |
| اللغة: |
English |
| بيانات الدورية: |
Publisher: American Chemical Society Country of Publication: United States NLM ID: 0370623 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1520-4995 (Electronic) Linking ISSN: 00062960 NLM ISO Abbreviation: Biochemistry Subsets: MEDLINE |
| أسماء مطبوعة: |
Original Publication: Washington, American Chemical Society. |
| مواضيع طبية MeSH: |
Ubiquitin Thiolesterase*/chemistry , Ubiquitin Thiolesterase*/metabolism , Protein Conformation*, Protein Folding ; Protein Unfolding ; Models, Molecular ; Humans ; Ion Mobility Spectrometry/methods |
| مستخلص: |
Knotted proteins are fascinating natural biomolecules whose backbones entangle themselves in a knot. Their particular knotted configurations provide them with a wide range of topological features. However, their folding/unfolding mechanisms, stability, and function are poorly understood. In the present work, native trapped ion mobility spectrometry-mass spectrometry (TIMS-MS) was used for characterizing structural features of two model knotted proteins: a Gordian 5 2 knot ubiquitin C-terminal hydrolase (UCH) and a Stevedore 6 1 knot (α-haloacid dehalogenase, DehI). Experimental results showed structural transitions of UCH and DehI as a function of solution composition (0-50% MeOH) and temperature ( T ∼20-95 °C). An increase in the protein charge states and collision cross sections (∼2750-8750 Å 2 and ∼3250-15,385 Å 2 for UCH and DehI, respectively) with the solution organic content (OC) and temperature suggested a three-step unfolding pathway with at least four structural transitions. Results also showed that the integrity of the UCH knot core was more resistant to thermal unfolding when compared to DehI; however, both knot cores can be disrupted with the increase in the solution OC. Additional enzymatic digestion experiments using carboxypeptidase Y combined with molecular dynamics simulations showed that the knot core was preserved between Glu20 and Glu188 and Arg89 and His304 residues for UCH and DehI, respectively, where disruption of the knot core led to structural collapse followed by unfolding events. This work highlights the potential of solution OC and temperature studies combined with native TIMS-MS for the comprehensive characterization of knotted proteins to gain a better understanding of their structural transitions. |
| المشرفين على المادة: |
EC 3.4.19.12 (Ubiquitin Thiolesterase) |
| تواريخ الأحداث: |
Date Created: 20240827 Date Completed: 20240917 Latest Revision: 20240917 |
| رمز التحديث: |
20240917 |
| DOI: |
10.1021/acs.biochem.4c00218 |
| PMID: |
39189377 |
| قاعدة البيانات: |
MEDLINE |
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