Interdomain interactions rearrangements control the reaction steps of a thermostable DNA alkyltransferase
| Autor: | Maria Ciaramella, Riccardo Miggiano, Menico Rizzi, Mario Serpe, Anna Valenti, Mosè Rossi, Giuseppe Perugino, Alberto Massarotti, Franca Rossi, Castrese Morrone, Giovanni del Monaco |
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| Přispěvatelé: | Morrone, Castrese, Miggiano, Riccardo, Serpe, Mario, Massarotti, Alberto, Valenti, Anna, Del Monaco, Giovanni, Rossi, Mosè, Rossi, Franca, Rizzi, Menico, Perugino, Giuseppe, Ciaramella, Maria |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Alkylation DNA Repair Fluorescent-based assay ved/biology.organism_classification_rank.species Perugino G Rossi F Biochemistry Catalysi chemistry.chemical_compound Rossi M Valenti A Ciaramella M Morrone C Protein alkylation biology Chemistry Protein Stability Miggiano R Sulfolobus solfataricus Del Monaco G DNA-Binding Proteins Massarotti A lipids (amino acids peptides and proteins) Alkylation damage Alkyltransferase Alkylating Agents Alkyl and Aryl Transferase Protein Domain DNA repair DNA-Binding Protein Biophysics Catalysis 03 medical and health sciences Protein Domains parasitic diseases Rizzi M Molecular Biology Alkyl and Aryl Transferases 030102 biochemistry & molecular biology ved/biology Crystal structure Active site DNA Conformational change Alkylating Agent Serpe M 030104 developmental biology biology.protein Cysteine |
| Zdroj: | Biochimica et biophysica acta 1861 (2017): 86–96. doi:10.1016/j.bbagen.2016.10.020 info:cnr-pdr/source/autori:Morrone C, Miggiano R, Serpe M, Massarotti A, Valenti A, Del Monaco G, Rossi M, Rossi F, Rizzi M, Perugino G, Ciaramella M./titolo:Interdomain interactions rearrangements control the reaction steps of a thermostable DNA alkyltransferase./doi:10.1016%2Fj.bbagen.2016.10.020/rivista:Biochimica et biophysica acta (Print)/anno:2017/pagina_da:86/pagina_a:96/intervallo_pagine:86–96/volume:1861 |
| ISSN: | 0304-4165 |
| DOI: | 10.1016/j.bbagen.2016.10.020 |
| Popis: | Background Alkylated DNA-protein alkyltransferases (AGTs) are conserved proteins that repair alkylation damage in DNA by using a single-step mechanism leading to irreversible alkylation of the catalytic cysteine in the active site. Trans-alkylation induces inactivation and destabilization of the protein, both in vitro and in vivo, likely triggering conformational changes. A complete picture of structural rearrangements occurring during the reaction cycle is missing, despite considerable interest raised by the peculiarity of AGT reaction, and the contribution of a functional AGT in limiting the efficacy of chemotherapy with alkylating drugs. Methods As a model for AGTs we have used a thermostable ortholog from the archaeon Sulfolobus solfataricus (SsOGT), performing biochemical, structural, molecular dynamics and in silico analysis of ligand-free, DNA-bound and mutated versions of the protein. Results Conformational changes occurring during lesion recognition and after the reaction, allowed us to identify a novel interaction network contributing to SsOGT stability, which is perturbed when a bulky adduct between the catalytic cysteine and the alkyl group is formed, a mandatory step toward the permanent protein alkylation. Conclusions Our data highlighted conformational changes and perturbation of intramolecular interaction occurring during lesion recognition and catalysis, confirming our previous hypothesis that coordination between the N- and C-terminal domains of SsOGT is important for protein activity and stability. General significance A general model of structural rearrangements occurring during the reaction cycle of AGTs is proposed. If confirmed, this model might be a starting point to design strategies to modulate AGT activity in therapeutic settings. |
| Databáze: | OpenAIRE |
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