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The structure, function and properties of sirohaem decarboxylase – an enzyme with structural homology to a transcription factor family that is part of the alternative haem biosynthesis pathway

The structure, function and properties of sirohaem decarboxylase – an enzyme with structural homology to a transcription factor family that is part of the alternative haem biosynthesis pathway
The structure, function and properties of sirohaem decarboxylase – an enzyme with structural homology to a transcription factor family that is part of the alternative haem biosynthesis pathway
Some bacteria and archaea synthesize haem by an alternative pathway, which involves the sequestration of sirohaem as a metabolic intermediate rather than as a prosthetic group. Along this pathway the two acetic acid side-chains attached to C12 and C18 are decarboxylated by sirohaem decarboxylase, a heterodimeric enzyme composed of AhbA and AhbB, to give didecarboxysirohaem. Further modifications catalysed by two related radical SAM enzymes, AhbC and AhbD, transform didecarboxysirohaem into Fe-coproporphyrin III and haem respectively. The characterization of sirohaem decarboxylase is reported in molecular detail. Recombinant versions of Desulfovibrio desulfuricansDesulfovibrio vulgaris and Methanosarcina barkeri AhbA/B have been produced and their physical properties compared. The D. vulgaris and M. barkeri enzyme complexes both copurify with haem, whose redox state influences the activity of the latter. The kinetic parameters of the D. desulfuricans enzyme have been determined, the enzyme crystallized and its structure has been elucidated. The topology of the enzyme reveals that it shares a structural similarity to the AsnC/Lrp family of transcription factors. The active site is formed in the cavity between the two subunits and a AhbA/B-product complex with didecarboxysirohaem has been obtained. A mechanism for the decarboxylation of the kinetically stable carboxyl groups is proposed.
0950-382X
247-261
Palmer, David J.
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Schroeder, Susanne
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Lawrence, Andrew D.
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Deery, Evelyne
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Lobo, Susana A.
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Saraiva, Ligia M.
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McLean, Kirsty J.
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Munro, Andrew W.
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Ferguson, Stuart J.
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Pickersgill, Richard W.
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Brown, David G.
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Warren, Martin J.
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et al.
Palmer, David J.
bdda8924-0580-40ae-b9b6-a59560596d70
Schroeder, Susanne
3bc51fbe-9fa2-4aec-b946-f2d5760a1129
Lawrence, Andrew D.
ce503b40-0155-486f-bb1d-26830b61b5f1
Deery, Evelyne
16c10c8d-1383-4ce6-95dd-dfcfa0168875
Lobo, Susana A.
6966e881-1121-4978-972f-18f191970239
Saraiva, Ligia M.
e47dd217-9095-42b3-91e3-2248d66586ee
McLean, Kirsty J.
1cf105ff-b3d1-4eb0-86ed-e39e94c56a0a
Munro, Andrew W.
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Ferguson, Stuart J.
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Pickersgill, Richard W.
9e186a55-43cd-4e87-bb1b-3b4bba7a3a6d
Brown, David G.
b8c25aa9-1bd7-4138-a637-0ad4e1608adc
Warren, Martin J.
048e9c28-25b0-4115-abc7-61d88ff4435b

Palmer, David J., Schroeder, Susanne and Lawrence, Andrew D. , et al. (2014) The structure, function and properties of sirohaem decarboxylase – an enzyme with structural homology to a transcription factor family that is part of the alternative haem biosynthesis pathway. Molecular Microbiology, 93 (2), 247-261. (doi:10.1111/mmi.12656).

Record type: Article

Abstract

Some bacteria and archaea synthesize haem by an alternative pathway, which involves the sequestration of sirohaem as a metabolic intermediate rather than as a prosthetic group. Along this pathway the two acetic acid side-chains attached to C12 and C18 are decarboxylated by sirohaem decarboxylase, a heterodimeric enzyme composed of AhbA and AhbB, to give didecarboxysirohaem. Further modifications catalysed by two related radical SAM enzymes, AhbC and AhbD, transform didecarboxysirohaem into Fe-coproporphyrin III and haem respectively. The characterization of sirohaem decarboxylase is reported in molecular detail. Recombinant versions of Desulfovibrio desulfuricansDesulfovibrio vulgaris and Methanosarcina barkeri AhbA/B have been produced and their physical properties compared. The D. vulgaris and M. barkeri enzyme complexes both copurify with haem, whose redox state influences the activity of the latter. The kinetic parameters of the D. desulfuricans enzyme have been determined, the enzyme crystallized and its structure has been elucidated. The topology of the enzyme reveals that it shares a structural similarity to the AsnC/Lrp family of transcription factors. The active site is formed in the cavity between the two subunits and a AhbA/B-product complex with didecarboxysirohaem has been obtained. A mechanism for the decarboxylation of the kinetically stable carboxyl groups is proposed.

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Accepted/In Press date: 24 May 2014
e-pub ahead of print date: 28 May 2014
Published date: 18 June 2014

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Local EPrints ID: 488492
URI: http://eprints.soton.ac.uk/id/eprint/488492
ISSN: 0950-382X
PURE UUID: a7237980-c97e-4aa7-b424-a825bd8d30f7
ORCID for Andrew D. Lawrence: ORCID iD orcid.org/0000-0002-5853-5409

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Date deposited: 22 Mar 2024 18:57
Last modified: 23 Mar 2024 03:10

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Contributors

Author: David J. Palmer
Author: Susanne Schroeder
Author: Andrew D. Lawrence ORCID iD
Author: Evelyne Deery
Author: Susana A. Lobo
Author: Ligia M. Saraiva
Author: Kirsty J. McLean
Author: Andrew W. Munro
Author: Stuart J. Ferguson
Author: Richard W. Pickersgill
Author: David G. Brown
Author: Martin J. Warren
Corporate Author: et al.

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