التفاصيل البيبلوغرافية
| العنوان: |
Computational Protein Stabilization Can Affect Folding Energy Landscapes and Lead to Domain-Swapped Dimers |
| المؤلفون: |
Klara Markova (10027446), Antonin Kunka (9457109), Klaudia Chmelova (10027449), Martin Havlasek (10027451), Petra Babkova (10027452), Sérgio M. Marques (9584429), Michal Vasina (9247186), Joan Planas-Iglesias (9247181), Radka Chaloupková (9247191), David Bednar (9247193), Zbynek Prokop (9705845), Jiri Damborsky (9242771), Martin Marek (6558650) |
| سنة النشر: |
2021 |
| المجموعة: |
Smithsonian Institution: Digital Repository |
| مصطلحات موضوعية: |
Biocatalysis, protein folding, protein design, α/β-hydrolase, haloalkane dehalogenase, domain-swapping, dimerization, energy landscape, oligomerization |
| الوصف: |
The functionality of a protein depends on its unique three-dimensional structure, which is a result of the folding process when the nascent polypeptide follows a funnel-like energy landscape to reach a global energy minimum. Computer-encoded algorithms are increasingly employed to stabilize native proteins for use in research and biotechnology applications. Here, we reveal a unique example where the computational stabilization of a monomeric α/β-hydrolase enzyme ( T m = 73.5°C; Δ T m > 23°C) affected the protein folding energy landscape. Introduction of eleven single-point stabilizing mutations based on force field calculations and evolutionary analysis yielded catalytically active domain-swapped intermediates trapped in local energy minima. Crystallographic structures revealed that these stabilizing mutations target cryptic hinge regions and newly introduced secondary interfaces, where they make extensive non-covalent interactions between the intertwined misfolded protomers. The existence of domain-swapped dimers in a solution is further confirmed experimentally by data obtained from SAXS and crosslinking mass spectrometry. Unfolding experiments showed that the domain-swapped dimers can be irreversibly converted into native-like monomers, suggesting that the domain-swapping occurs exclusively in vivo . Our findings uncovered hidden protein-folding consequences of computational protein design, which need to be taken into account when applying a rational stabilization to proteins of biological and pharmaceutical interest. |
| نوع الوثيقة: |
report |
| اللغة: |
unknown |
| Relation: |
https://figshare.com/articles/preprint/Computational_Protein_Stabilization_Can_Affect_Folding_Energy_Landscapes_and_Lead_to_Domain-Swapped_Dimers/13634021 |
| DOI: |
10.26434/chemrxiv.13634021.v1 |
| الاتاحة: |
https://doi.org/10.26434/chemrxiv.13634021.v1 |
| Rights: |
CC BY-NC-ND 4.0 |
| رقم الانضمام: |
edsbas.C652C455 |
| قاعدة البيانات: |
BASE |