Disruption of the dimerization interface of the sensing domain in the dimeric heme-based oxygen sensor AfGcHK abolishes bacterial signal transduction

0522236 - BTÚ 2021 RIV US eng J - Článek v odborném periodiku
Skálová, Tereza - Lengalová, A. - Dohnálek, Jan - Harlos, K. - Mihalcin, P. - Kolenko, Petr - Stranava, M. - Blaha, J. - Shimizu, T. - Martínková, M.
Disruption of the dimerization interface of the sensing domain in the dimeric heme-based oxygen sensor AfGcHK abolishes bacterial signal transduction.
Journal of Biological Chemistry. Roč. 295, č. 6 (2019), s. 1587-1597. ISSN 0021-9258. E-ISSN 1083-351X
Institucionální podpora: RVO:86652036
Klíčová slova: bacterial protein kinase * cell signaling * crystal structure * dimerization interface
Obor OECD: Biochemistry and molecular biology
Impakt faktor: 4.238, rok: 2019
Způsob publikování: Open access
https://www.sciencedirect.com/science/article/pii/S0021925817498589?via%3Dihub

The heme-based oxygen sensor protein AfGcHK is a globin-coupled histidine kinase in the soil bacterium Anaeromyxobacter sp. Fw109-5. Its C-terminal functional domain exhibits autophosphorylation activity induced by oxygen binding to the heme-Fe(II) complex located in the oxygen-sensing N-terminal globin domain. A detailed understanding of the signal transduction mechanisms in heme-containing sensor proteins remains elusive. Here, we investigated the role of the globin domain's dimerization interface in signal transduction in AfGcHK. We present a crystal structure of a monomeric imidazole-bound AfGcHK globin domain at 1.8 Å resolution, revealing that the helices of the WT globin dimer are under tension and suggesting that Tyr-15 plays a role in both this tension and the globin domain's dimerization. Biophysical experiments revealed that whereas the isolated WT globin domain is dimeric in solution, the Y15A and Y15G variants in which Tyr-15 is replaced with Ala or Gly, respectively, are monomeric. Additionally, we found that although the dimerization of the full-length protein is preserved via the kinase domain dimerization interface in all variants, full-length AfGcHK variants bearing the Y15A or Y15G substitutions lack enzymatic activity. The combined structural and biophysical results presented here indicate that Tyr-15 plays a key role in the dimerization of the globin domain of AfGcHK and that globin domain dimerization is essential for internal signal transduction and autophosphorylation in this protein. These findings provide critical insights into the signal transduction mechanism of the histidine kinase AfGcHK from Anaeromyxobacter.
Trvalý link: http://hdl.handle.net/11104/0306774