Bioluminescence Resonance Energy Transfer (BRET) Allows Monitoring the Barnase-Barstar Complex In Vivo
Bioluminescence resonance energy transfer (BRET) seems to be a promising biophysical technique to study protein–protein interactions within living cells due to a very specific reaction of bioluminescence that essentially decreases the background of other cellular components and light-induced destruc...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-02-01
|
| Series: | Biophysica |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2673-4125/2/1/7 |
| _version_ | 1797446956147015680 |
|---|---|
| author | Victor Marchenkov Tanya Ivashina Natalia Marchenko Vladimir Ksenzenko Gennady Semisotnov |
| author_facet | Victor Marchenkov Tanya Ivashina Natalia Marchenko Vladimir Ksenzenko Gennady Semisotnov |
| author_sort | Victor Marchenkov |
| collection | DOAJ |
| description | Bioluminescence resonance energy transfer (BRET) seems to be a promising biophysical technique to study protein–protein interactions within living cells due to a very specific reaction of bioluminescence that essentially decreases the background of other cellular components and light-induced destruction of biomacromolecules. An important direction of the development of this technique is the study of known strong protein–protein complexes in vivo and the estimation of an average distance between chromophores of the donor and acceptor. Here, we demonstrate an in vivo interaction between barnase fused with luciferase (from <i>Renilla reniformis,</i> RLuc) and barstar fused with EGFP (enhanced green fluorescent protein of <i>Aequorea victoria</i>) monitored by BRET. The distance between the luciferase and EGFP chromophores within the complex has been evaluated as equal to (56 ± 2) Å. |
| first_indexed | 2024-03-09T13:48:57Z |
| format | Article |
| id | doaj.art-485dd50cd84a49ef8a3805f0be6eab97 |
| institution | Directory Open Access Journal |
| issn | 2673-4125 |
| language | English |
| last_indexed | 2024-03-09T13:48:57Z |
| publishDate | 2022-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biophysica |
| spelling | doaj.art-485dd50cd84a49ef8a3805f0be6eab972023-11-30T20:53:48ZengMDPI AGBiophysica2673-41252022-02-0121727810.3390/biophysica2010007Bioluminescence Resonance Energy Transfer (BRET) Allows Monitoring the Barnase-Barstar Complex In VivoVictor Marchenkov0Tanya Ivashina1Natalia Marchenko2Vladimir Ksenzenko3Gennady Semisotnov4Institute of Protein Research, Russian Academy of Sciences, Institutskaya Street 4, 142290 Pushchino, RussiaSkryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Prospect Nauki 5, 142290 Pushchino, RussiaInstitute of Protein Research, Russian Academy of Sciences, Institutskaya Street 4, 142290 Pushchino, RussiaInstitute of Protein Research, Russian Academy of Sciences, Institutskaya Street 4, 142290 Pushchino, RussiaInstitute of Protein Research, Russian Academy of Sciences, Institutskaya Street 4, 142290 Pushchino, RussiaBioluminescence resonance energy transfer (BRET) seems to be a promising biophysical technique to study protein–protein interactions within living cells due to a very specific reaction of bioluminescence that essentially decreases the background of other cellular components and light-induced destruction of biomacromolecules. An important direction of the development of this technique is the study of known strong protein–protein complexes in vivo and the estimation of an average distance between chromophores of the donor and acceptor. Here, we demonstrate an in vivo interaction between barnase fused with luciferase (from <i>Renilla reniformis,</i> RLuc) and barstar fused with EGFP (enhanced green fluorescent protein of <i>Aequorea victoria</i>) monitored by BRET. The distance between the luciferase and EGFP chromophores within the complex has been evaluated as equal to (56 ± 2) Å.https://www.mdpi.com/2673-4125/2/1/7protein–protein interactionsfusion proteinsphotoproteinsBRETbarnasebarstar |
| spellingShingle | Victor Marchenkov Tanya Ivashina Natalia Marchenko Vladimir Ksenzenko Gennady Semisotnov Bioluminescence Resonance Energy Transfer (BRET) Allows Monitoring the Barnase-Barstar Complex In Vivo Biophysica protein–protein interactions fusion proteins photoproteins BRET barnase barstar |
| title | Bioluminescence Resonance Energy Transfer (BRET) Allows Monitoring the Barnase-Barstar Complex In Vivo |
| title_full | Bioluminescence Resonance Energy Transfer (BRET) Allows Monitoring the Barnase-Barstar Complex In Vivo |
| title_fullStr | Bioluminescence Resonance Energy Transfer (BRET) Allows Monitoring the Barnase-Barstar Complex In Vivo |
| title_full_unstemmed | Bioluminescence Resonance Energy Transfer (BRET) Allows Monitoring the Barnase-Barstar Complex In Vivo |
| title_short | Bioluminescence Resonance Energy Transfer (BRET) Allows Monitoring the Barnase-Barstar Complex In Vivo |
| title_sort | bioluminescence resonance energy transfer bret allows monitoring the barnase barstar complex in vivo |
| topic | protein–protein interactions fusion proteins photoproteins BRET barnase barstar |
| url | https://www.mdpi.com/2673-4125/2/1/7 |
| work_keys_str_mv | AT victormarchenkov bioluminescenceresonanceenergytransferbretallowsmonitoringthebarnasebarstarcomplexinvivo AT tanyaivashina bioluminescenceresonanceenergytransferbretallowsmonitoringthebarnasebarstarcomplexinvivo AT nataliamarchenko bioluminescenceresonanceenergytransferbretallowsmonitoringthebarnasebarstarcomplexinvivo AT vladimirksenzenko bioluminescenceresonanceenergytransferbretallowsmonitoringthebarnasebarstarcomplexinvivo AT gennadysemisotnov bioluminescenceresonanceenergytransferbretallowsmonitoringthebarnasebarstarcomplexinvivo |