Structure of the effector-binding domain of the arabinose repressor AraR from Bacillus subtilis

Kateřina Procházková, Kateřina Čermáková, Petr Pachl, Irena Sieglová, Milan Fábry, Zbyszek Otwinowski, Pavlína Řezáčová

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

In Bacillus subtilis, the arabinose repressor AraR negatively controls the expression of genes in the metabolic pathway of arabinose-containing polysaccharides. The protein is composed of two domains of different phylogenetic origin and function: an N-terminal DNA-binding domain belonging to the GntR family and a C-terminal effector-binding domain that shows similarity to members of the GalR/LacI family. The crystal structure of the C-terminal effector-binding domain of AraR in complex with the effector L-arabinose has been determined at 2.2 Å resolution. The L-arabinose binding affinity was characterized by isothermal titration calorimetry and differential scanning fluorimetry; the K d value was 8.4 ± 0.4 μM. The effect of L-arabinose on the protein oligomeric state was investigated in solution and detailed analysis of the crystal identified a dimer organization which is distinctive from that of other members of the GalR/LacI family.

Original languageEnglish (US)
Pages (from-to)176-185
Number of pages10
JournalActa Crystallographica Section D: Biological Crystallography
Volume68
Issue number2
DOIs
StatePublished - Feb 2012

Fingerprint

Arabinose
Bacillus subtilis
Fluorometry
Differential Scanning Calorimetry
Metabolic Networks and Pathways
Polysaccharides
Proteins
Gene Expression
DNA

Keywords

  • differential scanning fluorimetry
  • dimeric interface
  • dimerization
  • effector binding
  • isothermal titration calorimetry
  • repressors

ASJC Scopus subject areas

  • Structural Biology

Cite this

Structure of the effector-binding domain of the arabinose repressor AraR from Bacillus subtilis. / Procházková, Kateřina; Čermáková, Kateřina; Pachl, Petr; Sieglová, Irena; Fábry, Milan; Otwinowski, Zbyszek; Řezáčová, Pavlína.

In: Acta Crystallographica Section D: Biological Crystallography, Vol. 68, No. 2, 02.2012, p. 176-185.

Research output: Contribution to journalArticle

Procházková, Kateřina ; Čermáková, Kateřina ; Pachl, Petr ; Sieglová, Irena ; Fábry, Milan ; Otwinowski, Zbyszek ; Řezáčová, Pavlína. / Structure of the effector-binding domain of the arabinose repressor AraR from Bacillus subtilis. In: Acta Crystallographica Section D: Biological Crystallography. 2012 ; Vol. 68, No. 2. pp. 176-185.
@article{b52388fb7ef447ba8be5671dcd89deb9,
title = "Structure of the effector-binding domain of the arabinose repressor AraR from Bacillus subtilis",
abstract = "In Bacillus subtilis, the arabinose repressor AraR negatively controls the expression of genes in the metabolic pathway of arabinose-containing polysaccharides. The protein is composed of two domains of different phylogenetic origin and function: an N-terminal DNA-binding domain belonging to the GntR family and a C-terminal effector-binding domain that shows similarity to members of the GalR/LacI family. The crystal structure of the C-terminal effector-binding domain of AraR in complex with the effector L-arabinose has been determined at 2.2 {\AA} resolution. The L-arabinose binding affinity was characterized by isothermal titration calorimetry and differential scanning fluorimetry; the K d value was 8.4 ± 0.4 μM. The effect of L-arabinose on the protein oligomeric state was investigated in solution and detailed analysis of the crystal identified a dimer organization which is distinctive from that of other members of the GalR/LacI family.",
keywords = "differential scanning fluorimetry, dimeric interface, dimerization, effector binding, isothermal titration calorimetry, repressors",
author = "Kateřina Proch{\'a}zkov{\'a} and Kateřina Čerm{\'a}kov{\'a} and Petr Pachl and Irena Sieglov{\'a} and Milan F{\'a}bry and Zbyszek Otwinowski and Pavl{\'i}na Řez{\'a}čov{\'a}",
year = "2012",
month = "2",
doi = "10.1107/S090744491105414X",
language = "English (US)",
volume = "68",
pages = "176--185",
journal = "Acta Crystallographica Section D: Structural Biology",
issn = "0907-4449",
publisher = "John Wiley and Sons Inc.",
number = "2",

}

TY - JOUR

T1 - Structure of the effector-binding domain of the arabinose repressor AraR from Bacillus subtilis

AU - Procházková, Kateřina

AU - Čermáková, Kateřina

AU - Pachl, Petr

AU - Sieglová, Irena

AU - Fábry, Milan

AU - Otwinowski, Zbyszek

AU - Řezáčová, Pavlína

PY - 2012/2

Y1 - 2012/2

N2 - In Bacillus subtilis, the arabinose repressor AraR negatively controls the expression of genes in the metabolic pathway of arabinose-containing polysaccharides. The protein is composed of two domains of different phylogenetic origin and function: an N-terminal DNA-binding domain belonging to the GntR family and a C-terminal effector-binding domain that shows similarity to members of the GalR/LacI family. The crystal structure of the C-terminal effector-binding domain of AraR in complex with the effector L-arabinose has been determined at 2.2 Å resolution. The L-arabinose binding affinity was characterized by isothermal titration calorimetry and differential scanning fluorimetry; the K d value was 8.4 ± 0.4 μM. The effect of L-arabinose on the protein oligomeric state was investigated in solution and detailed analysis of the crystal identified a dimer organization which is distinctive from that of other members of the GalR/LacI family.

AB - In Bacillus subtilis, the arabinose repressor AraR negatively controls the expression of genes in the metabolic pathway of arabinose-containing polysaccharides. The protein is composed of two domains of different phylogenetic origin and function: an N-terminal DNA-binding domain belonging to the GntR family and a C-terminal effector-binding domain that shows similarity to members of the GalR/LacI family. The crystal structure of the C-terminal effector-binding domain of AraR in complex with the effector L-arabinose has been determined at 2.2 Å resolution. The L-arabinose binding affinity was characterized by isothermal titration calorimetry and differential scanning fluorimetry; the K d value was 8.4 ± 0.4 μM. The effect of L-arabinose on the protein oligomeric state was investigated in solution and detailed analysis of the crystal identified a dimer organization which is distinctive from that of other members of the GalR/LacI family.

KW - differential scanning fluorimetry

KW - dimeric interface

KW - dimerization

KW - effector binding

KW - isothermal titration calorimetry

KW - repressors

UR - http://www.scopus.com/inward/record.url?scp=84856488349&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84856488349&partnerID=8YFLogxK

U2 - 10.1107/S090744491105414X

DO - 10.1107/S090744491105414X

M3 - Article

C2 - 22281747

AN - SCOPUS:84856488349

VL - 68

SP - 176

EP - 185

JO - Acta Crystallographica Section D: Structural Biology

JF - Acta Crystallographica Section D: Structural Biology

SN - 0907-4449

IS - 2

ER -