A novel c-di-GMP binding domain in glycosyltransferase BgsA is responsible for the synthesis of a mixed-linkage β-glucan
Abstract BgsA is the glycosyltransferase (GT) involved in the synthesis of a linear mixed-linkage β-glucan (MLG), a recently described exopolysaccharide activated by c-di-GMP in Sinorhizobium meliloti and other Rhizobiales. Although BgsA displays sequence and structural homology with bacterial cellu...
Guardado en:
| Autores principales: | , , , , , |
|---|---|
| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
Nature Portfolio
2017
|
| Materias: | |
| Acceso en línea: | https://doaj.org/article/99701fe68fec4ef8a2e19b448ca9722f |
| Etiquetas: |
Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
|
| id |
oai:doaj.org-article:99701fe68fec4ef8a2e19b448ca9722f |
|---|---|
| record_format |
dspace |
| spelling |
oai:doaj.org-article:99701fe68fec4ef8a2e19b448ca9722f2021-12-02T12:30:25ZA novel c-di-GMP binding domain in glycosyltransferase BgsA is responsible for the synthesis of a mixed-linkage β-glucan10.1038/s41598-017-09290-22045-2322https://doaj.org/article/99701fe68fec4ef8a2e19b448ca9722f2017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-09290-2https://doaj.org/toc/2045-2322Abstract BgsA is the glycosyltransferase (GT) involved in the synthesis of a linear mixed-linkage β-glucan (MLG), a recently described exopolysaccharide activated by c-di-GMP in Sinorhizobium meliloti and other Rhizobiales. Although BgsA displays sequence and structural homology with bacterial cellulose synthases (CS), it does not contain any predictable c-di-GMP binding domain. In this work we demonstrate that the cytoplasmic C-terminal domain of BgsA (C-BgsA) binds c-di-GMP with both high affinity (KD = 0.23 μM) and specificity. C-BgsA is structurally different to the otherwise equivalent cytoplasmic C-terminal domain of CS, and does not contain PilZ motifs for c-di-GMP recognition. A combination of random and site-directed mutagenesis with surface plasmon resonance (SPR) allowed identification of the C-BgsA residues which are important not only for c-di-GMP binding, but also for BgsA GT activity. The results suggest that the C-BgsA domain is important for both, c-di-GMP binding and GT activity of BgsA. In contrast to bacterial CS where c-di-GMP has been proposed as a derepressor of GT activity, we hypothesize that the C-terminal domain of BgsA plays an active role in BgsA GT activity upon binding c-di-GMP.Daniel Pérez-MendozaDaniela BertinettiRobin LorenzMaría-Trinidad GallegosFriedrich W. HerbergJuan SanjuánNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017) |
| institution |
DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
Medicine R Science Q |
| spellingShingle |
Medicine R Science Q Daniel Pérez-Mendoza Daniela Bertinetti Robin Lorenz María-Trinidad Gallegos Friedrich W. Herberg Juan Sanjuán A novel c-di-GMP binding domain in glycosyltransferase BgsA is responsible for the synthesis of a mixed-linkage β-glucan |
| description |
Abstract BgsA is the glycosyltransferase (GT) involved in the synthesis of a linear mixed-linkage β-glucan (MLG), a recently described exopolysaccharide activated by c-di-GMP in Sinorhizobium meliloti and other Rhizobiales. Although BgsA displays sequence and structural homology with bacterial cellulose synthases (CS), it does not contain any predictable c-di-GMP binding domain. In this work we demonstrate that the cytoplasmic C-terminal domain of BgsA (C-BgsA) binds c-di-GMP with both high affinity (KD = 0.23 μM) and specificity. C-BgsA is structurally different to the otherwise equivalent cytoplasmic C-terminal domain of CS, and does not contain PilZ motifs for c-di-GMP recognition. A combination of random and site-directed mutagenesis with surface plasmon resonance (SPR) allowed identification of the C-BgsA residues which are important not only for c-di-GMP binding, but also for BgsA GT activity. The results suggest that the C-BgsA domain is important for both, c-di-GMP binding and GT activity of BgsA. In contrast to bacterial CS where c-di-GMP has been proposed as a derepressor of GT activity, we hypothesize that the C-terminal domain of BgsA plays an active role in BgsA GT activity upon binding c-di-GMP. |
| format |
article |
| author |
Daniel Pérez-Mendoza Daniela Bertinetti Robin Lorenz María-Trinidad Gallegos Friedrich W. Herberg Juan Sanjuán |
| author_facet |
Daniel Pérez-Mendoza Daniela Bertinetti Robin Lorenz María-Trinidad Gallegos Friedrich W. Herberg Juan Sanjuán |
| author_sort |
Daniel Pérez-Mendoza |
| title |
A novel c-di-GMP binding domain in glycosyltransferase BgsA is responsible for the synthesis of a mixed-linkage β-glucan |
| title_short |
A novel c-di-GMP binding domain in glycosyltransferase BgsA is responsible for the synthesis of a mixed-linkage β-glucan |
| title_full |
A novel c-di-GMP binding domain in glycosyltransferase BgsA is responsible for the synthesis of a mixed-linkage β-glucan |
| title_fullStr |
A novel c-di-GMP binding domain in glycosyltransferase BgsA is responsible for the synthesis of a mixed-linkage β-glucan |
| title_full_unstemmed |
A novel c-di-GMP binding domain in glycosyltransferase BgsA is responsible for the synthesis of a mixed-linkage β-glucan |
| title_sort |
novel c-di-gmp binding domain in glycosyltransferase bgsa is responsible for the synthesis of a mixed-linkage β-glucan |
| publisher |
Nature Portfolio |
| publishDate |
2017 |
| url |
https://doaj.org/article/99701fe68fec4ef8a2e19b448ca9722f |
| work_keys_str_mv |
AT danielperezmendoza anovelcdigmpbindingdomaininglycosyltransferasebgsaisresponsibleforthesynthesisofamixedlinkagebglucan AT danielabertinetti anovelcdigmpbindingdomaininglycosyltransferasebgsaisresponsibleforthesynthesisofamixedlinkagebglucan AT robinlorenz anovelcdigmpbindingdomaininglycosyltransferasebgsaisresponsibleforthesynthesisofamixedlinkagebglucan AT mariatrinidadgallegos anovelcdigmpbindingdomaininglycosyltransferasebgsaisresponsibleforthesynthesisofamixedlinkagebglucan AT friedrichwherberg anovelcdigmpbindingdomaininglycosyltransferasebgsaisresponsibleforthesynthesisofamixedlinkagebglucan AT juansanjuan anovelcdigmpbindingdomaininglycosyltransferasebgsaisresponsibleforthesynthesisofamixedlinkagebglucan AT danielperezmendoza novelcdigmpbindingdomaininglycosyltransferasebgsaisresponsibleforthesynthesisofamixedlinkagebglucan AT danielabertinetti novelcdigmpbindingdomaininglycosyltransferasebgsaisresponsibleforthesynthesisofamixedlinkagebglucan AT robinlorenz novelcdigmpbindingdomaininglycosyltransferasebgsaisresponsibleforthesynthesisofamixedlinkagebglucan AT mariatrinidadgallegos novelcdigmpbindingdomaininglycosyltransferasebgsaisresponsibleforthesynthesisofamixedlinkagebglucan AT friedrichwherberg novelcdigmpbindingdomaininglycosyltransferasebgsaisresponsibleforthesynthesisofamixedlinkagebglucan AT juansanjuan novelcdigmpbindingdomaininglycosyltransferasebgsaisresponsibleforthesynthesisofamixedlinkagebglucan |
| _version_ |
1718394411394531328 |