Crystal structure of Mycobacterium tuberculosis FadB2 implicated in mycobacterial β-oxidation

Jonathan A.G. Cox, Rebecca C. Taylor, Alistair K. Brown, Samuel Attoe, Gurdyal S. Besra, Klaus Fütterer*

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

The intracellular pathogen Mycobacterium tuberculosis is the causative agent of tuberculosis, which is a leading cause of mortality worldwide. The survival of M. tuberculosis in host macrophages through long-lasting periods of persistence depends, in part, on breaking down host cell lipids as a carbon source. The critical role of fatty-acid catabolism in this organism is underscored by the extensive redundancy of the genes implicated in β-oxidation (∼100 genes). In a previous study, the enzymology of the M. tuberculosisl-3-hydroxyacyl-CoA dehydrogenase FadB2 was characterized. Here, the crystal structure of this enzyme in a ligand-free form is reported at 2.1 Å resolution. FadB2 crystallized as a dimer with three unique dimer copies per asymmetric unit. The structure of the monomer reveals a dual Rossmann-fold motif in the N-terminal domain, while the helical C-terminal domain mediates dimer formation. Comparison with the CoA- and NAD + -bound human orthologue mitochondrial hydroxyacyl-CoA dehydrogenase shows extensive conservation of the residues that mediate substrate and cofactor binding. Superposition with the multi-catalytic homologue M. tuberculosis FadB, which forms a trifunctional complex with the thiolase FadA, indicates that FadB has developed structural features that prevent its self-association as a dimer. Conversely, FadB2 is unable to substitute for FadB in the tetrameric FadA–FadB complex as it lacks the N-terminal hydratase domain of FadB. Instead, FadB2 may functionally (or physically) associate with the enoyl-CoA hydratase EchA8 and the thiolases FadA2, FadA3, FadA4 or FadA6 as suggested by interrogation of the STRING protein-network database.

Original languageEnglish
Pages (from-to)101-108
Number of pages8
JournalActa Crystallographica Section D: Structural Biology
Volume75
Issue numberPt 1
DOIs
Publication statusPublished - 1 Jan 2019

Fingerprint

Coenzyme A
Mycobacterium tuberculosis
Tuberculosis
Enoyl-CoA Hydratase
3-Hydroxyacyl-CoA Dehydrogenase
Hydro-Lyases
Protein Databases
NAD
Genes
Oxidoreductases
Fatty Acids
Carbon
Macrophages
Ligands
Lipids
Survival
Mortality
Enzymes

Bibliographical note

© 2019 The Authors. Published under an open-access Creative Commons Attribution (CC-BY) Licence. Acta Cryst. (2019). D75, 101-108

Funding: This work is supported by funding from the Medical Research
Council (MRC; MR/S000542/1).

Keywords

  • l-3-hydroxyacyl-CoA dehydrogenase
  • mycobacterial β-oxidation
  • Mycobacterium tuberculosis
  • X-ray crystallography
  • 3-Hydroxyacyl CoA Dehydrogenases/chemistry
  • Enoyl-CoA Hydratase/metabolism
  • Oxidation-Reduction
  • Humans
  • Protein Multimerization
  • Protein Binding
  • Crystallography, X-Ray
  • Mycobacterium tuberculosis/enzymology

Cite this

Cox, J. A. G., Taylor, R. C., Brown, A. K., Attoe, S., Besra, G. S., & Fütterer, K. (2019). Crystal structure of Mycobacterium tuberculosis FadB2 implicated in mycobacterial β-oxidation. Acta Crystallographica Section D: Structural Biology, 75(Pt 1), 101-108. https://doi.org/10.1107/S2059798318017242
Cox, Jonathan A.G. ; Taylor, Rebecca C. ; Brown, Alistair K. ; Attoe, Samuel ; Besra, Gurdyal S. ; Fütterer, Klaus. / Crystal structure of Mycobacterium tuberculosis FadB2 implicated in mycobacterial β-oxidation. In: Acta Crystallographica Section D: Structural Biology. 2019 ; Vol. 75, No. Pt 1. pp. 101-108.
@article{a8a6c30164a1441196a7e0bfd2f178b9,
title = "Crystal structure of Mycobacterium tuberculosis FadB2 implicated in mycobacterial β-oxidation",
abstract = "The intracellular pathogen Mycobacterium tuberculosis is the causative agent of tuberculosis, which is a leading cause of mortality worldwide. The survival of M. tuberculosis in host macrophages through long-lasting periods of persistence depends, in part, on breaking down host cell lipids as a carbon source. The critical role of fatty-acid catabolism in this organism is underscored by the extensive redundancy of the genes implicated in β-oxidation (∼100 genes). In a previous study, the enzymology of the M. tuberculosisl-3-hydroxyacyl-CoA dehydrogenase FadB2 was characterized. Here, the crystal structure of this enzyme in a ligand-free form is reported at 2.1 {\AA} resolution. FadB2 crystallized as a dimer with three unique dimer copies per asymmetric unit. The structure of the monomer reveals a dual Rossmann-fold motif in the N-terminal domain, while the helical C-terminal domain mediates dimer formation. Comparison with the CoA- and NAD + -bound human orthologue mitochondrial hydroxyacyl-CoA dehydrogenase shows extensive conservation of the residues that mediate substrate and cofactor binding. Superposition with the multi-catalytic homologue M. tuberculosis FadB, which forms a trifunctional complex with the thiolase FadA, indicates that FadB has developed structural features that prevent its self-association as a dimer. Conversely, FadB2 is unable to substitute for FadB in the tetrameric FadA–FadB complex as it lacks the N-terminal hydratase domain of FadB. Instead, FadB2 may functionally (or physically) associate with the enoyl-CoA hydratase EchA8 and the thiolases FadA2, FadA3, FadA4 or FadA6 as suggested by interrogation of the STRING protein-network database.",
keywords = "l-3-hydroxyacyl-CoA dehydrogenase, mycobacterial β-oxidation, Mycobacterium tuberculosis, X-ray crystallography, 3-Hydroxyacyl CoA Dehydrogenases/chemistry, Enoyl-CoA Hydratase/metabolism, Oxidation-Reduction, Humans, Protein Multimerization, Protein Binding, Crystallography, X-Ray, Mycobacterium tuberculosis/enzymology",
author = "Cox, {Jonathan A.G.} and Taylor, {Rebecca C.} and Brown, {Alistair K.} and Samuel Attoe and Besra, {Gurdyal S.} and Klaus F{\"u}tterer",
note = "{\circledC} 2019 The Authors. Published under an open-access Creative Commons Attribution (CC-BY) Licence. Acta Cryst. (2019). D75, 101-108 Funding: This work is supported by funding from the Medical Research Council (MRC; MR/S000542/1).",
year = "2019",
month = "1",
day = "1",
doi = "10.1107/S2059798318017242",
language = "English",
volume = "75",
pages = "101--108",
number = "Pt 1",

}

Cox, JAG, Taylor, RC, Brown, AK, Attoe, S, Besra, GS & Fütterer, K 2019, 'Crystal structure of Mycobacterium tuberculosis FadB2 implicated in mycobacterial β-oxidation', Acta Crystallographica Section D: Structural Biology, vol. 75, no. Pt 1, pp. 101-108. https://doi.org/10.1107/S2059798318017242

Crystal structure of Mycobacterium tuberculosis FadB2 implicated in mycobacterial β-oxidation. / Cox, Jonathan A.G.; Taylor, Rebecca C.; Brown, Alistair K.; Attoe, Samuel; Besra, Gurdyal S.; Fütterer, Klaus.

In: Acta Crystallographica Section D: Structural Biology, Vol. 75, No. Pt 1, 01.01.2019, p. 101-108.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Crystal structure of Mycobacterium tuberculosis FadB2 implicated in mycobacterial β-oxidation

AU - Cox, Jonathan A.G.

AU - Taylor, Rebecca C.

AU - Brown, Alistair K.

AU - Attoe, Samuel

AU - Besra, Gurdyal S.

AU - Fütterer, Klaus

N1 - © 2019 The Authors. Published under an open-access Creative Commons Attribution (CC-BY) Licence. Acta Cryst. (2019). D75, 101-108 Funding: This work is supported by funding from the Medical Research Council (MRC; MR/S000542/1).

PY - 2019/1/1

Y1 - 2019/1/1

N2 - The intracellular pathogen Mycobacterium tuberculosis is the causative agent of tuberculosis, which is a leading cause of mortality worldwide. The survival of M. tuberculosis in host macrophages through long-lasting periods of persistence depends, in part, on breaking down host cell lipids as a carbon source. The critical role of fatty-acid catabolism in this organism is underscored by the extensive redundancy of the genes implicated in β-oxidation (∼100 genes). In a previous study, the enzymology of the M. tuberculosisl-3-hydroxyacyl-CoA dehydrogenase FadB2 was characterized. Here, the crystal structure of this enzyme in a ligand-free form is reported at 2.1 Å resolution. FadB2 crystallized as a dimer with three unique dimer copies per asymmetric unit. The structure of the monomer reveals a dual Rossmann-fold motif in the N-terminal domain, while the helical C-terminal domain mediates dimer formation. Comparison with the CoA- and NAD + -bound human orthologue mitochondrial hydroxyacyl-CoA dehydrogenase shows extensive conservation of the residues that mediate substrate and cofactor binding. Superposition with the multi-catalytic homologue M. tuberculosis FadB, which forms a trifunctional complex with the thiolase FadA, indicates that FadB has developed structural features that prevent its self-association as a dimer. Conversely, FadB2 is unable to substitute for FadB in the tetrameric FadA–FadB complex as it lacks the N-terminal hydratase domain of FadB. Instead, FadB2 may functionally (or physically) associate with the enoyl-CoA hydratase EchA8 and the thiolases FadA2, FadA3, FadA4 or FadA6 as suggested by interrogation of the STRING protein-network database.

AB - The intracellular pathogen Mycobacterium tuberculosis is the causative agent of tuberculosis, which is a leading cause of mortality worldwide. The survival of M. tuberculosis in host macrophages through long-lasting periods of persistence depends, in part, on breaking down host cell lipids as a carbon source. The critical role of fatty-acid catabolism in this organism is underscored by the extensive redundancy of the genes implicated in β-oxidation (∼100 genes). In a previous study, the enzymology of the M. tuberculosisl-3-hydroxyacyl-CoA dehydrogenase FadB2 was characterized. Here, the crystal structure of this enzyme in a ligand-free form is reported at 2.1 Å resolution. FadB2 crystallized as a dimer with three unique dimer copies per asymmetric unit. The structure of the monomer reveals a dual Rossmann-fold motif in the N-terminal domain, while the helical C-terminal domain mediates dimer formation. Comparison with the CoA- and NAD + -bound human orthologue mitochondrial hydroxyacyl-CoA dehydrogenase shows extensive conservation of the residues that mediate substrate and cofactor binding. Superposition with the multi-catalytic homologue M. tuberculosis FadB, which forms a trifunctional complex with the thiolase FadA, indicates that FadB has developed structural features that prevent its self-association as a dimer. Conversely, FadB2 is unable to substitute for FadB in the tetrameric FadA–FadB complex as it lacks the N-terminal hydratase domain of FadB. Instead, FadB2 may functionally (or physically) associate with the enoyl-CoA hydratase EchA8 and the thiolases FadA2, FadA3, FadA4 or FadA6 as suggested by interrogation of the STRING protein-network database.

KW - l-3-hydroxyacyl-CoA dehydrogenase

KW - mycobacterial β-oxidation

KW - Mycobacterium tuberculosis

KW - X-ray crystallography

KW - 3-Hydroxyacyl CoA Dehydrogenases/chemistry

KW - Enoyl-CoA Hydratase/metabolism

KW - Oxidation-Reduction

KW - Humans

KW - Protein Multimerization

KW - Protein Binding

KW - Crystallography, X-Ray

KW - Mycobacterium tuberculosis/enzymology

UR - http://www.scopus.com/inward/record.url?scp=85060028616&partnerID=8YFLogxK

UR - http://10.1107/S2059798318017242/mn5119sup1.pdf

U2 - 10.1107/S2059798318017242

DO - 10.1107/S2059798318017242

M3 - Article

C2 - 30644849

VL - 75

SP - 101

EP - 108

IS - Pt 1

ER -

Cox JAG, Taylor RC, Brown AK, Attoe S, Besra GS, Fütterer K. Crystal structure of Mycobacterium tuberculosis FadB2 implicated in mycobacterial β-oxidation. Acta Crystallographica Section D: Structural Biology. 2019 Jan 1;75(Pt 1):101-108. https://doi.org/10.1107/S2059798318017242