Mutations of Asp540 and the domain-connecting residues synergistically enhance Pyrococcus furiosus DNA ligase activity
| Autor: | Hirokazu Nishida, Maiko Tanabe, Sonoko Ishino, Yoshizumi Ishino |
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| Rok vydání: | 2013 |
| Předmět: |
Thermostable enzyme
Models Molecular DNA Ligases Molecular Sequence Data Biophysics DNA ligase activity Biology medicine.disease_cause Biochemistry Protein Structure Secondary chemistry.chemical_compound Structural Biology Genetics medicine Humans Amino Acid Sequence Molecular Biology Adenylylation chemistry.chemical_classification Aspartic Acid DNA ligase Mutation Cell Biology Mutational analysis biology.organism_classification Archaea Protein Structure Tertiary Pyrococcus furiosus Amino Acid Substitution chemistry Helix Protein engineering DNA |
| Zdroj: | FEBS Letters. 588:230-235 |
| ISSN: | 0014-5793 |
| DOI: | 10.1016/j.febslet.2013.10.037 |
| Popis: | The structure of Pyrococcus furiosus DNA ligase (PfuLig), which architecturally resembles human DNA ligase I (hLigI), revealed that the C-terminal helix stabilizes the closed conformation through several ionic interactions between two domains (adenylylation domain (AdD) and C-terminal OB-fold domain (OBD)). This helix is oriented differently in DNA-bound hLigI, suggesting that the disruption of its interactions with AdD facilitates DNA binding. Previously, we demonstrated that the replacement of Asp540 with arginine improves the ligation activity. Here we report that the combination of the Asp540-replacement and the elimination of ionic residues in the helix, forming interactions with AdD, effectively enhanced the activity. |
| Databáze: | OpenAIRE |
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