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[FeFe]-hydrogenase maturation: HydG-catalyzed synthesis of carbon monoxide

[FeFe]-hydrogenase maturation: HydG-catalyzed synthesis of carbon monoxide
[FeFe]-hydrogenase maturation: HydG-catalyzed synthesis of carbon monoxide
Biosynthesis of the unusual organometallic H-cluster at the active site of the [FeFe]-hydrogenase requires three accessory proteins, two of which are radical AdoMet enzymes (HydE, HydG) and one of which is a GTPase (HydF). We demonstrate here that HydG catalyzes the synthesis of CO using tyrosine as a substrate. CO production was detected by using deoxyhemoglobin as a reporter and monitoring the appearance of the characteristic visible spectroscopic features of carboxyhemoglobin. Assays utilizing 13C-tyrosine were analyzed by FTIR to confirm the production of HbCO and to demonstrate that the CO product was synthesized from tyrosine. CO ligation is a common feature at the active sites of the [FeFe], [NiFe], and [Fe]-only hydrogenases; however, this is the first report of the enzymatic synthesis of CO in hydrogenase maturation.
0002-7863
9247-9249
Shepard, Eric M.
2f916a0e-62bf-410b-bf52-b0756e86365e
Duffus, Benjamin R.
3eb25fae-1b53-4bf3-a266-775f936592a9
George, Simon J.
20847a69-0612-4765-8874-932d5e17082c
McGlynn, Shawn E.
894be6e2-ef61-43f4-b3fa-077afd7735cb
Challand, Martin R.
03bb1ded-9274-4525-8377-8e5930f30d65
Swanson, Kevin D.
ef502c53-f150-4daf-aedd-ac5765a4df32
Roach, Peter L.
ca94060c-4443-482b-af3e-979243488ba9
Cramer, Stephen P.
f5bba2d6-c816-44a8-ac6f-a011371feae3
Peters, John W.
84377e0f-e6d8-4e56-aed8-584ec85c28ad
Broderick, Joan B.
9eb68505-070d-4ba2-a2ad-71c942330a1a
Shepard, Eric M.
2f916a0e-62bf-410b-bf52-b0756e86365e
Duffus, Benjamin R.
3eb25fae-1b53-4bf3-a266-775f936592a9
George, Simon J.
20847a69-0612-4765-8874-932d5e17082c
McGlynn, Shawn E.
894be6e2-ef61-43f4-b3fa-077afd7735cb
Challand, Martin R.
03bb1ded-9274-4525-8377-8e5930f30d65
Swanson, Kevin D.
ef502c53-f150-4daf-aedd-ac5765a4df32
Roach, Peter L.
ca94060c-4443-482b-af3e-979243488ba9
Cramer, Stephen P.
f5bba2d6-c816-44a8-ac6f-a011371feae3
Peters, John W.
84377e0f-e6d8-4e56-aed8-584ec85c28ad
Broderick, Joan B.
9eb68505-070d-4ba2-a2ad-71c942330a1a

Shepard, Eric M., Duffus, Benjamin R., George, Simon J., McGlynn, Shawn E., Challand, Martin R., Swanson, Kevin D., Roach, Peter L., Cramer, Stephen P., Peters, John W. and Broderick, Joan B. (2010) [FeFe]-hydrogenase maturation: HydG-catalyzed synthesis of carbon monoxide. Journal of the American Chemical Society, 132 (27), 9247-9249. (doi:10.1021/ja1012273). (PMID:20565074)

Record type: Article

Abstract

Biosynthesis of the unusual organometallic H-cluster at the active site of the [FeFe]-hydrogenase requires three accessory proteins, two of which are radical AdoMet enzymes (HydE, HydG) and one of which is a GTPase (HydF). We demonstrate here that HydG catalyzes the synthesis of CO using tyrosine as a substrate. CO production was detected by using deoxyhemoglobin as a reporter and monitoring the appearance of the characteristic visible spectroscopic features of carboxyhemoglobin. Assays utilizing 13C-tyrosine were analyzed by FTIR to confirm the production of HbCO and to demonstrate that the CO product was synthesized from tyrosine. CO ligation is a common feature at the active sites of the [FeFe], [NiFe], and [Fe]-only hydrogenases; however, this is the first report of the enzymatic synthesis of CO in hydrogenase maturation.

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Published date: 21 June 2010
Organisations: Chemistry

Identifiers

Local EPrints ID: 177025
URI: http://eprints.soton.ac.uk/id/eprint/177025
ISSN: 0002-7863
PURE UUID: 2b48467b-ee35-4cd3-9fae-6164e403f35e
ORCID for Peter L. Roach: ORCID iD orcid.org/0000-0001-9880-2877

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Date deposited: 28 Mar 2011 14:10
Last modified: 17 Dec 2019 01:52

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Contributors

Author: Eric M. Shepard
Author: Benjamin R. Duffus
Author: Simon J. George
Author: Shawn E. McGlynn
Author: Martin R. Challand
Author: Kevin D. Swanson
Author: Peter L. Roach ORCID iD
Author: Stephen P. Cramer
Author: John W. Peters
Author: Joan B. Broderick

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