Isolation and Characterization of Monomeric Human RAD51 : A Novel Tool for Investigating Homologous Recombination in Cancer
© 2023 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH..
DNA repair protein RAD51 is a key player in the homologous recombination pathway. Upon DNA damage, RAD51 is transported into the nucleus by BRCA2, where it can repair DNA double-strand breaks. Due to the structural complexity and dynamics, researchers have not yet clarified the mechanistic details of every step of RAD51 recruitment and DNA repair. RAD51 possesses an intrinsic tendency to form oligomeric structures, which make it challenging to conduct biochemical and biophysical investigations. Here, for the first time, we report on the isolation and characterization of a human monomeric RAD51 recombinant form, obtained through a double mutation, which preserves the protein's integrity and functionality. We investigated different buffers to identify the most suitable condition needed to definitively stabilize the monomer. The monomer of human RAD51 provides the community with a unique biological tool for investigating RAD51-mediated homologous recombination, and paves the way for more reliable structural, mechanistic, and drug discovery studies.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2023 |
|---|---|
| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:62 |
|---|---|
| Enthalten in: |
Angewandte Chemie (International ed. in English) - 62(2023), 51 vom: 18. Dez., Seite e202312517 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Rinaldi, Francesco [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
DNA Damage |
|---|
| Anmerkungen: |
Date Completed 12.01.2024 Date Revised 12.01.2024 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1002/anie.202312517 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM364160810 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM364160810 | ||
| 003 | DE-627 | ||
| 005 | 20240114234249.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231226s2023 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1002/anie.202312517 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1257.xml |
| 035 | |a (DE-627)NLM364160810 | ||
| 035 | |a (NLM)37924230 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Rinaldi, Francesco |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Isolation and Characterization of Monomeric Human RAD51 |b A Novel Tool for Investigating Homologous Recombination in Cancer |
| 264 | 1 | |c 2023 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 12.01.2024 | ||
| 500 | |a Date Revised 12.01.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2023 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH. | ||
| 520 | |a DNA repair protein RAD51 is a key player in the homologous recombination pathway. Upon DNA damage, RAD51 is transported into the nucleus by BRCA2, where it can repair DNA double-strand breaks. Due to the structural complexity and dynamics, researchers have not yet clarified the mechanistic details of every step of RAD51 recruitment and DNA repair. RAD51 possesses an intrinsic tendency to form oligomeric structures, which make it challenging to conduct biochemical and biophysical investigations. Here, for the first time, we report on the isolation and characterization of a human monomeric RAD51 recombinant form, obtained through a double mutation, which preserves the protein's integrity and functionality. We investigated different buffers to identify the most suitable condition needed to definitively stabilize the monomer. The monomer of human RAD51 provides the community with a unique biological tool for investigating RAD51-mediated homologous recombination, and paves the way for more reliable structural, mechanistic, and drug discovery studies | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a DNA Damage | |
| 650 | 4 | |a Protein Engineering | |
| 650 | 4 | |a Protein-Protein Interactions | |
| 650 | 4 | |a RAD51 | |
| 650 | 4 | |a Structural Biology | |
| 650 | 7 | |a RAD51 protein, human |2 NLM | |
| 650 | 7 | |a EC 2.7.7.- |2 NLM | |
| 650 | 7 | |a Rad51 Recombinase |2 NLM | |
| 650 | 7 | |a EC 2.7.7.- |2 NLM | |
| 650 | 7 | |a Recombinant Proteins |2 NLM | |
| 700 | 1 | |a Schipani, Fabrizio |e verfasserin |4 aut | |
| 700 | 1 | |a Balboni, Beatrice |e verfasserin |4 aut | |
| 700 | 1 | |a Catalano, Federico |e verfasserin |4 aut | |
| 700 | 1 | |a Marotta, Roberto |e verfasserin |4 aut | |
| 700 | 1 | |a Myers, Samuel H |e verfasserin |4 aut | |
| 700 | 1 | |a Previtali, Viola |e verfasserin |4 aut | |
| 700 | 1 | |a Veronesi, Marina |e verfasserin |4 aut | |
| 700 | 1 | |a Scietti, Luigi |e verfasserin |4 aut | |
| 700 | 1 | |a Cecatiello, Valentina |e verfasserin |4 aut | |
| 700 | 1 | |a Pasqualato, Sebastiano |e verfasserin |4 aut | |
| 700 | 1 | |a Ortega, Jose Antonio |e verfasserin |4 aut | |
| 700 | 1 | |a Girotto, Stefania |e verfasserin |4 aut | |
| 700 | 1 | |a Cavalli, Andrea |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Angewandte Chemie (International ed. in English) |d 1964 |g 62(2023), 51 vom: 18. Dez., Seite e202312517 |w (DE-627)NLM000105422 |x 1521-3773 |7 nnns |
| 773 | 1 | 8 | |g volume:62 |g year:2023 |g number:51 |g day:18 |g month:12 |g pages:e202312517 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1002/anie.202312517 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 62 |j 2023 |e 51 |b 18 |c 12 |h e202312517 | ||