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https://hdl.handle.net/123456789/1216| Τύπος: | Αναρτημένη ανακοίνωση (poster) |
| Τίτλος: | Segmental isotope labelling of the multidomain La protein for NMR structural study |
| Συγγραφέας: | [EL] Αργυρίου, Αικατερίνη[EN] Argyriou, Aikaterini [EL] Μαχαλιώτης, Γεώργιος[EN] Machaliotis, Georgios [EN] von Schroetter, Christine [EL] Καλιάτση, Ελένη[EN] Kaliatsi, Eleni [FR] Allain, Fréd́eric Hai Trieu [EL] Σταθόπουλος, Κωνσταντίνος[EN] Stathopoulos, Constantinos [EL] Σπυρούλιας, Γεώργιος[EN] Spyroulias, Georgios |
| Ημερομηνία: | 03/03/2021 |
| Περίληψη: | La protein plays a key-role in processing of primary transcripts including tRNAs, by facilitating their proper folding. Although La seems to recognize and bind the 3’ poly(U) tails of many artificial small RNAs, this feature is under debate for the natural ligands of La and the exact mechanism is still unknown. Human La contains four distinct domains, namely La motif (LaM), two RNA recognition motifs (RRM1 and RRM2α) and a variable C-terminus region. Until today the structure of the full-length La protein and the role of RRM2α remain elusive. Determination of the solution NMR structure of the full-length La protein (408 a.a.) with the conventional labeling techniques is a difficult task due to the size. Among the non-standard protocols that can provide solutions, segmental isotopic labeling has been identified as a powerful tool for NMR structural studies of large multi-domain proteins like La protein. This technique allows the selective observation of protein segment(s) within a protein, simplifying thus the spectral data for large proteins. Therefore, we used segmental labeling techniques to elucidate the structure and RNAbinding properties of each La domain, while being part of the whole protein. Each construct was prepared for expression in fusion with an intein tag. Expressed Protein Ligation technique (EPL) was based on the reaction of a C-terminal α-thioester of RRM1 with the N-terminal Cys195 of RRM2α. The reaction results in the formation of a native peptide bond after cleavage of intein and purification on a Chitin column. The 15N-TROSY spectrum of {15N-RRM1}-{14N-RRM2α} and {14N-RRM1}-{15N-RRM2α} revealed that RRM1 and RRM2 expressed with the linker between them are well folded and match fairly well with reference spectra of individual 15N-RRM1 and 15N-RRM2α, respectively |
| Γλώσσα: | Αγγλικά |
| Σελίδες: | 6 |
| Θεματική κατηγορία: | [EL] Βιοχημεία και Μοριακή βιολογία[EN] Biochemistry and Molecular Biology |
| Κάτοχος πνευματικών δικαιωμάτων: | © The Author(s) 2021 |
| Όνομα εκδήλωσης: | Διαδικτυακό Συνέδριο Ελληνικής Εταιρείας Βιοχημείας & Μοριακής Βιολογίας “Chemical, structural biology and disease treatment” |
| Τοποθεσία εκδήλωσης: | Virtual Conference |
| Ημ/νία έναρξης εκδήλωσης: | 03/03/2021 |
| Ημ/νία λήξης εκδήλωσης: | 03/03/2021 |
| Σημειώσεις: | Conference Program : https://eebmb.gr/images/synedria/PROGRAM_3_Mar_2021.pdf Co-financed by Greece and the European Union |
| Εμφανίζεται στις συλλογές: | Μεταδιδακτορικοί ερευνητές |
Αρχεία σε αυτό το τεκμήριο:
| Αρχείο | Περιγραφή | Σελίδες | Μέγεθος | Μορφότυπος | Έκδοση | Άδεια | |
|---|---|---|---|---|---|---|---|
| argyriou eebmb 2021.pdf | 6 διαφάνειες σελίδες | 531.26 kB | Adobe PDF | - |  | Δείτε/ανοίξτε |