Aims: Structural and functional characterization of the globin-coupled sensors (GCSs) from Azotobacter vinelandii (AvGReg) and Bordetella pertussis (BpeGReg). Results: Ultraviolet/visible and resonance Raman spectroscopies confirm the presence in AvGReg and BpeGReg of a globin domain capable of reversible gaseous ligand binding. In AvGReg, an influence of the transmitter domain on the heme proximal region of the globin domain can be seen, and k′CO is higher than for other GCSs. The O2 binding kinetics suggests the presence of an open and a closed conformation. As for BpeGReg, the fully oxygenated AvGReg show a very high diguanylate cyclase activity. The carbon monoxide rebinding to BpeGReg indicates that intra- and intermolecular interactions influence the ligand binding. The globin domains of both proteins (AvGReg globin domain and BpeGRegGb with cysteines (Cys16, 45, 114, 154) mutated to serines [BpeGReg-Gb∗]) share the same GCS fold, a similar proximal but a different distal side structure. They homodimerize through a G-H helical bundle as in other GCSs. However, BpeGReg-Gb∗ shows also a second dimerization mode. Innovation: This article extends our knowledge on the GCS proteins and contributes to a better understanding of the GCSs role in the formation of bacterial biofilms. Conclusions: AvGReg and BpeGReg conform to the GCS family, share a similar overall structure, but they have different properties in terms of the ligand binding. In particular, AvGReg shows an open and a closed conformation that in the latter form will very tightly bind oxygen. BpeGReg has only one closed conformation. In both proteins, it is the fully oxygenated GCS form that catalyzes the production of the second messenger.
Structural and Functional Characterization of the Globin-Coupled Sensors of Azotobacter vinelandii and Bordetella pertussis / F. Germani, M. Nardini, A. De Schutter, B. Cuypers, H. Berghmans, M.-. Van Hauwaert, S. Bruno, A. Mozzarelli, L. Moens, S. Van Doorslaer, M. Bolognesi, A. Pesce, S. Dewilde. - In: ANTIOXIDANTS & REDOX SIGNALING. - ISSN 1523-0864. - 32:6(2020), pp. 378-395.
|Titolo:||Structural and Functional Characterization of the Globin-Coupled Sensors of Azotobacter vinelandii and Bordetella pertussis|
|Parole Chiave:||heme-based sensor; oxygen affinity; c-di-GMP and enzyme specificity; biofilm; crystal structure|
|Settore Scientifico Disciplinare:||Settore BIO/10 - Biochimica|
Settore FIS/07 - Fisica Applicata(Beni Culturali, Ambientali, Biol.e Medicin)
|Data di pubblicazione:||2020|
|Data ahead of print / Data di stampa:||3-dic-2019|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1089/ars.2018.7690|
|Appare nelle tipologie:||01 - Articolo su periodico|
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