Solution structure of the SGTA dimerisation domain and investigation of its interactions with the ubiquitin-like domains of BAG6 and UBL4A.
The BAG6 complex resides in the cytosol and acts as a sorting point to target diverse hydrophobic protein substrates along their appropriate paths, including proteasomal degradation and ER membrane insertion. Composed of a trimeric complex of BAG6, TRC35 and UBL4A, the BAG6 complex is closely associ...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4240585?pdf=render |
| _version_ | 1818538700025364480 |
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| author | John F Darby Ewelina M Krysztofinska Peter J Simpson Aline C Simon Pawel Leznicki Newran Sriskandarajah David S Bishop Lisa R Hale Caterina Alfano Maria R Conte Santiago Martínez-Lumbreras Arjun Thapaliya Stephen High Rivka L Isaacson |
| author_facet | John F Darby Ewelina M Krysztofinska Peter J Simpson Aline C Simon Pawel Leznicki Newran Sriskandarajah David S Bishop Lisa R Hale Caterina Alfano Maria R Conte Santiago Martínez-Lumbreras Arjun Thapaliya Stephen High Rivka L Isaacson |
| author_sort | John F Darby |
| collection | DOAJ |
| description | The BAG6 complex resides in the cytosol and acts as a sorting point to target diverse hydrophobic protein substrates along their appropriate paths, including proteasomal degradation and ER membrane insertion. Composed of a trimeric complex of BAG6, TRC35 and UBL4A, the BAG6 complex is closely associated with SGTA, a co-chaperone from which it can obtain hydrophobic substrates.SGTA consists of an N-terminal dimerisation domain (SGTA_NT), a central tetratricopeptide repeat (TPR) domain, and a glutamine rich region towards the C-terminus. Here we solve a solution structure of the SGTA dimerisation domain and use biophysical techniques to investigate its interaction with two different UBL domains from the BAG6 complex. The SGTA_NT structure is a dimer with a tight hydrophobic interface connecting two sets of four alpha helices. Using a combination of NMR chemical shift perturbation, isothermal titration calorimetry (ITC) and microscale thermophoresis (MST) experiments we have biochemically characterised the interactions of SGTA with components of the BAG6 complex, the ubiquitin-like domain (UBL) containing proteins UBL4A and BAG6. We demonstrate that the UBL domains from UBL4A and BAG6 directly compete for binding to SGTA at the same site. Using a combination of structural and interaction data we have implemented the HADDOCK protein-protein interaction docking tool to generate models of the SGTA-UBL complexes.This atomic level information contributes to our understanding of the way in which hydrophobic proteins have their fate decided by the collaboration between SGTA and the BAG6 complex. |
| first_indexed | 2024-12-11T21:32:20Z |
| format | Article |
| id | doaj.art-8e8a952f4dc54536822cf9d134d1aff8 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-11T21:32:20Z |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-8e8a952f4dc54536822cf9d134d1aff82022-12-22T00:50:08ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-01911e11328110.1371/journal.pone.0113281Solution structure of the SGTA dimerisation domain and investigation of its interactions with the ubiquitin-like domains of BAG6 and UBL4A.John F DarbyEwelina M KrysztofinskaPeter J SimpsonAline C SimonPawel LeznickiNewran SriskandarajahDavid S BishopLisa R HaleCaterina AlfanoMaria R ConteSantiago Martínez-LumbrerasArjun ThapaliyaStephen HighRivka L IsaacsonThe BAG6 complex resides in the cytosol and acts as a sorting point to target diverse hydrophobic protein substrates along their appropriate paths, including proteasomal degradation and ER membrane insertion. Composed of a trimeric complex of BAG6, TRC35 and UBL4A, the BAG6 complex is closely associated with SGTA, a co-chaperone from which it can obtain hydrophobic substrates.SGTA consists of an N-terminal dimerisation domain (SGTA_NT), a central tetratricopeptide repeat (TPR) domain, and a glutamine rich region towards the C-terminus. Here we solve a solution structure of the SGTA dimerisation domain and use biophysical techniques to investigate its interaction with two different UBL domains from the BAG6 complex. The SGTA_NT structure is a dimer with a tight hydrophobic interface connecting two sets of four alpha helices. Using a combination of NMR chemical shift perturbation, isothermal titration calorimetry (ITC) and microscale thermophoresis (MST) experiments we have biochemically characterised the interactions of SGTA with components of the BAG6 complex, the ubiquitin-like domain (UBL) containing proteins UBL4A and BAG6. We demonstrate that the UBL domains from UBL4A and BAG6 directly compete for binding to SGTA at the same site. Using a combination of structural and interaction data we have implemented the HADDOCK protein-protein interaction docking tool to generate models of the SGTA-UBL complexes.This atomic level information contributes to our understanding of the way in which hydrophobic proteins have their fate decided by the collaboration between SGTA and the BAG6 complex.http://europepmc.org/articles/PMC4240585?pdf=render |
| spellingShingle | John F Darby Ewelina M Krysztofinska Peter J Simpson Aline C Simon Pawel Leznicki Newran Sriskandarajah David S Bishop Lisa R Hale Caterina Alfano Maria R Conte Santiago Martínez-Lumbreras Arjun Thapaliya Stephen High Rivka L Isaacson Solution structure of the SGTA dimerisation domain and investigation of its interactions with the ubiquitin-like domains of BAG6 and UBL4A. PLoS ONE |
| title | Solution structure of the SGTA dimerisation domain and investigation of its interactions with the ubiquitin-like domains of BAG6 and UBL4A. |
| title_full | Solution structure of the SGTA dimerisation domain and investigation of its interactions with the ubiquitin-like domains of BAG6 and UBL4A. |
| title_fullStr | Solution structure of the SGTA dimerisation domain and investigation of its interactions with the ubiquitin-like domains of BAG6 and UBL4A. |
| title_full_unstemmed | Solution structure of the SGTA dimerisation domain and investigation of its interactions with the ubiquitin-like domains of BAG6 and UBL4A. |
| title_short | Solution structure of the SGTA dimerisation domain and investigation of its interactions with the ubiquitin-like domains of BAG6 and UBL4A. |
| title_sort | solution structure of the sgta dimerisation domain and investigation of its interactions with the ubiquitin like domains of bag6 and ubl4a |
| url | http://europepmc.org/articles/PMC4240585?pdf=render |
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