• Je něco špatně v tomto záznamu ?

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

T. Skalova, A. Lengalova, J. Dohnalek, K. Harlos, P. Mihalcin, P. Kolenko, M. Stranava, J. Blaha, T. Shimizu, M. Martínková,

. 2020 ; 295 (6) : 1587-1597. [pub] 20191230

Jazyk angličtina Země Spojené státy americké

Typ dokumentu časopisecké články, práce podpořená grantem

Perzistentní odkaz   https://www.medvik.cz/link/bmc20028512

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.

Citace poskytuje Crossref.org

000      
00000naa a2200000 a 4500
001      
bmc20028512
003      
CZ-PrNML
005      
20210114154112.0
007      
ta
008      
210105s2020 xxu f 000 0|eng||
009      
AR
024    7_
$a 10.1074/jbc.RA119.011574 $2 doi
035    __
$a (PubMed)31914416
040    __
$a ABA008 $b cze $d ABA008 $e AACR2
041    0_
$a eng
044    __
$a xxu
100    1_
$a Skalova, Tereza $u Institute of Biotechnology of the Czech Academy of Sciences, v.v.i., Biocev, Vestec, 252 50 Czech Republic.
245    10
$a Disruption of the dimerization interface of the sensing domain in the dimeric heme-based oxygen sensor AfGcHK abolishes bacterial signal transduction / $c T. Skalova, A. Lengalova, J. Dohnalek, K. Harlos, P. Mihalcin, P. Kolenko, M. Stranava, J. Blaha, T. Shimizu, M. Martínková,
520    9_
$a 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.
650    _2
$a bakteriální proteiny $x chemie $x metabolismus $7 D001426
650    _2
$a krystalografie rentgenová $7 D018360
650    _2
$a globiny $x chemie $x metabolismus $7 D005914
650    _2
$a histidinkinasa $x chemie $x metabolismus $7 D000071677
650    _2
$a molekulární modely $7 D008958
650    _2
$a Myxococcales $x chemie $x metabolismus $7 D009231
650    _2
$a fosforylace $7 D010766
650    _2
$a konformace proteinů $7 D011487
650    _2
$a konformace proteinů, alfa-helix $7 D000072756
650    _2
$a proteinové domény $7 D000072417
650    _2
$a multimerizace proteinu $7 D055503
650    _2
$a signální transdukce $7 D015398
655    _2
$a časopisecké články $7 D016428
655    _2
$a práce podpořená grantem $7 D013485
700    1_
$a Lengalova, Alzbeta $u Department of Biochemistry, Faculty of Science, Charles University, Prague 2, 128 43 Czech Republic.
700    1_
$a Dohnalek, Jan $u Institute of Biotechnology of the Czech Academy of Sciences, v.v.i., Biocev, Vestec, 252 50 Czech Republic.
700    1_
$a Harlos, Karl $u Division of Structural Biology, The Wellcome Trust Centre for Human Genetics, University of Oxford, OX3 7BN Oxford, United Kingdom.
700    1_
$a Mihalcin, Peter $u Department of Biochemistry, Faculty of Science, Charles University, Prague 2, 128 43 Czech Republic.
700    1_
$a Kolenko, Petr $u Institute of Biotechnology of the Czech Academy of Sciences, v.v.i., Biocev, Vestec, 252 50 Czech Republic. FNSPE, Czech Technical University in Prague, Brehova 7, Prague 1, 115 19 Czech Republic.
700    1_
$a Stranava, Martin $u Department of Biochemistry, Faculty of Science, Charles University, Prague 2, 128 43 Czech Republic.
700    1_
$a Blaha, Jan $u Department of Biochemistry, Faculty of Science, Charles University, Prague 2, 128 43 Czech Republic.
700    1_
$a Shimizu, Toru $u Department of Biochemistry, Faculty of Science, Charles University, Prague 2, 128 43 Czech Republic.
700    1_
$a Martínková, Markéta $u Department of Biochemistry, Faculty of Science, Charles University, Prague 2, 128 43 Czech Republic marketa.martinkova@natur.cuni.cz.
773    0_
$w MED00002546 $t The Journal of biological chemistry $x 1083-351X $g Roč. 295, č. 6 (2020), s. 1587-1597
856    41
$u https://pubmed.ncbi.nlm.nih.gov/31914416 $y Pubmed
910    __
$a ABA008 $b sig $c sign $y a $z 0
990    __
$a 20210105 $b ABA008
991    __
$a 20210114154109 $b ABA008
999    __
$a ok $b bmc $g 1608847 $s 1119692
BAS    __
$a 3
BAS    __
$a PreBMC
BMC    __
$a 2020 $b 295 $c 6 $d 1587-1597 $e 20191230 $i 1083-351X $m The Journal of biological chemistry $n J Biol Chem $x MED00002546
LZP    __
$a Pubmed-20210105
Zpět

Najít záznam

Citační ukazatele

Možnosti archivace