The FOXO forkhead transcription factors are potent transcriptional activators involved in a wide range of key biological processes. In this work, the real-time kinetics of the interaction between the FOXO4-DNA binding domain (FOXO4-DBD) and the DNA was studied by using surface plasmon resonance (SPR). SPR analysis revealed that the interaction between FOXO4-DBD and the double stranded DNA containing either the insulin-responsive or the Daf-16 family member-binding element is preferably described by using a conformational change model which suggests a structural change of FOXO4-DBD upon binding to the DNA. This was further confirmed by using the time-resolved tryptophan fluorescence anisotropy decay measurements which revealed profound reduction of segmental dynamics of FOXO4-DBD upon the complex formation. Alanine scanning of amino acid residues engaged in polar contacts with the DNA showed that certain non-specific contacts with the DNA backbone are very important for both the binding affinity and the binding specificity of FOXO4-DBD.

Department of Physical and Macromolecular Chemistry Faculty of Science Charles University Prague 12843 Prague Czech Republic

000      

00000naa a2200000 a 4500

001      

bmc14074601

003      

CZ-PrNML

005      

20141007111747.0

007      

ta

008      

141006s2013 ne f 000 0|eng||

009      

AR

024    7_

$a 10.1016/j.bpc.2013.09.002 $2 doi

035    __

$a (PubMed)24121535

040    __

$a ABA008 $b cze $d ABA008 $e AACR2

041    0_

$a eng

044    __

$a ne

100    1_

$a Vacha, Petr $u Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University in Prague, 12843 Prague, Czech Republic.

245    10

$a Detailed kinetic analysis of the interaction between the FOXO4-DNA-binding domain and DNA / $c P. Vacha, I. Zuskova, L. Bumba, P. Herman, J. Vecer, V. Obsilova, T. Obsil,

520    9_

$a The FOXO forkhead transcription factors are potent transcriptional activators involved in a wide range of key biological processes. In this work, the real-time kinetics of the interaction between the FOXO4-DNA binding domain (FOXO4-DBD) and the DNA was studied by using surface plasmon resonance (SPR). SPR analysis revealed that the interaction between FOXO4-DBD and the double stranded DNA containing either the insulin-responsive or the Daf-16 family member-binding element is preferably described by using a conformational change model which suggests a structural change of FOXO4-DBD upon binding to the DNA. This was further confirmed by using the time-resolved tryptophan fluorescence anisotropy decay measurements which revealed profound reduction of segmental dynamics of FOXO4-DBD upon the complex formation. Alanine scanning of amino acid residues engaged in polar contacts with the DNA showed that certain non-specific contacts with the DNA backbone are very important for both the binding affinity and the binding specificity of FOXO4-DBD.

650    _2

$a vazebná místa $7 D001665

650    _2

$a DNA $x chemie $7 D004247

650    _2

$a lidé $7 D006801

650    _2

$a kinetika $7 D007700

650    _2

$a molekulární modely $7 D008958

650    _2

$a terciární struktura proteinů $7 D017434

650    _2

$a povrchová plasmonová rezonance $7 D020349

650    _2

$a transkripční faktory $x chemie $7 D014157

655    _2

$a časopisecké články $7 D016428

655    _2

$a práce podpořená grantem $7 D013485

700    1_

$a Zuskova, Iva

700    1_

$a Bumba, Ladislav

700    1_

$a Herman, Petr

700    1_

$a Vecer, Jaroslav

700    1_

$a Obsilova, Veronika

700    1_

$a Obsil, Tomas

773    0_

$w MED00000773 $t Biophysical chemistry $x 1873-4200 $g Roč. 184, č. - (2013), s. 68-78

856    41

$u https://pubmed.ncbi.nlm.nih.gov/24121535 $y Pubmed

910    __

$a ABA008 $b sig $c sign $y a $z 0

990    __

$a 20141006 $b ABA008

991    __

$a 20141007112225 $b ABA008

999    __

$a ok $b bmc $g 1042484 $s 873513

BAS    __

$a 3

BAS    __

$a PreBMC

BMC    __

$a 2013 $b 184 $c - $d 68-78 $i 1873-4200 $m Biophysical chemistry $n Biophys Chem $x MED00000773

LZP    __

$a Pubmed-20141006