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https://hdl.handle.net/123456789/888| Τύπος: | Άρθρο σε επιστημονικό περιοδικό |
| Τίτλος: | Consensus protein engineering on the thermostable histone‑like bacterial protein HUs significantly improves stability and DNA binding affinity |
| Συγγραφέας: | [EL] Γεωργούλης, Αναστάσιος[EN] Georgoulis, Anastasios [EL] Λουκά, Μαρία[EN] Louka, Maria [EL] Μυλωνάς, Στράτος[EN] Mylonas, Stratos [EL] Σταύρος, Φιλήμων[EN] Stavros, Philemon [EL] Νούνεσης, Γιώργος[EN] Nounesis, George [EL] Βοργιάς, Κωνσταντίνος[EN] Vorgias, Constantinos |
| Ημερομηνία: | 24/01/2020 |
| Περίληψη: | Consensus-based protein engineering strategy has been applied to various proteins and it can lead to the design of proteins with enhanced biological performance. Histone-like HUs comprise a protein family with sequence variety within a highly conserved 3D-fold. HU function includes compacting and regulating bacterial DNA in a wide range of biological conditions in bacteria. To explore the possible impact of consensus-based design in the thermodynamic stability of HU proteins, the approach was applied using a dataset of sequences derived from a group of 40 mesostable, thermostable, and hyperthermostable HUs. The consensus-derived HU protein was named HUBest, since it is expected to perform best. The synthetic HU gene was overexpressed in E. coli and the recombinant protein was purified. Subsequently, HUBest was characterized concerning its correct folding and thermodynamic stability, as well as its ability to interact with plasmid DNA. A substantial increase in HUBest stability at high temperatures is observed. HUBest has significantly improved biological performance at ambience temperature, presenting very low Kd values for binding plasmid DNA as indicated from the Gibbs energy profile of HUBest. This Kd may be associated to conformational changes leading to decreased thermodynamic stability and, therefore, higher flexibility at ambient temperature. |
| Γλώσσα: | Αγγλικά |
| Σελίδες: | 14 |
| DOI: | 10.1007/s00792-020-01154-4 |
| ISSN: | 1431-0651 |
| Θεματική κατηγορία: | [EL] Βιοχημεία και Μοριακή βιολογία[EN] Biochemistry and Molecular Biology |
| Λέξεις-κλειδιά: | HU histone-like protein; Consensus design; Thermostability; DNA binding activity |
| Κάτοχος πνευματικών δικαιωμάτων: | © Springer Japan KK, part of Springer Nature 2020 |
| Ηλεκτρονική διεύθυνση του τεκμηρίου στον εκδότη: | https://link.springer.com/article/10.1007/s00792-020-01154-4 |
| Ηλεκτρονική διεύθυνση περιοδικού: | https://www.springer.com/journal/792 |
| Τίτλος πηγής δημοσίευσης: | Extremophiles |
| Τόμος: | 24 |
| Σελίδες τεκμηρίου (στην πηγή): | 293–306 |
| Σημειώσεις: | We acknowledge technical support by the SPC facility at EMBL Hamburg in the frame of Biostruct-X (EE FP7). We would also like to acknowledge technical assistance from A. Tsoka and N Papandreou for biocomputing in our department. F. S. has been supported in part by the ARISTEIA I program (Grant Number 1125), administered by the General Secretariat of Research and Technology of Greece, co-financed by the European Social Fund and the State of Greece. The financial support of "The National Research Infrastructures on Integrated Structural Biology, Drug Screening Efforts and Drug target functional characterization” with the acronym “INSPIRED" and code (MIS) 5002550 from the Greek Government is also acknowledged |
| Εμφανίζεται στις συλλογές: | Μεταδιδακτορικοί ερευνητές |
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| Georgoulis et al. 2020.pdf | δημοσίευση σε επιστημονικό περιοδικό | 1.85 MB | Adobe PDF | - |  | Δείτε/ανοίξτε |