The Pseudomonas aeruginosa nirE gene encodes the S-adenosyl-L-methionine- dependent uroporphyrinogen III methyltransferase required for heme d1 biosynthesis

Storbeck, Sonja, Walther, Johannes, Müller, Judith, Parmar, Vina, Schiebel, Hans Martin, Kemken, Dorit, Dülcks, Thomas, Warren, Martin J. ORCID: https://orcid.org/0000-0002-6028-6456 and Layer, Gunhild (2009) The Pseudomonas aeruginosa nirE gene encodes the S-adenosyl-L-methionine- dependent uroporphyrinogen III methyltransferase required for heme d1 biosynthesis. FEBS Journal, 276 (20). pp. 5973-5982. ISSN 1742-464X

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Abstract

Biosynthesis of heme d1, the essential prosthetic group of the dissimilatory nitrite reductase cytochrome cd1, requires the methylation of the tetrapyrrole precursor uroporphyrinogen III at positions C-2 and C-7. We produced Pseudomonas aeruginosa NirE, a putative S-adenosyl-l-methionine (SAM)-dependent uroporphyrinogen III methyltransferase, as a recombinant protein in Escherichia coli and purified it to apparent homogeneity by metal chelate and gel filtration chromatography. Analytical gel filtration of purified NirE indicated that the recombinant protein is a homodimer. NirE was shown to be a SAM-dependent uroporphyrinogen III methyltransferase that catalyzes the conversion of uroporphyrinogen III into precorrin-2 in vivo and in vitro. A specific activity of 316.8 nmol of precorrin-2 h-1·mg-1 of NirE was found for the conversion of uroporphyrinogen III to precorrin-2. At high enzyme concentrations NirE catalyzed an overmethylation of uroporphyrinogen III, resulting in the formation of trimethylpyrrocorphin. Substrate inhibition was observed at uroporphyrinogen III concentrations above 17 m. The protein did bind SAM, although not with the same avidity as reported for other SAM-dependent uroporphyrinogen III methyltransferases involved in siroheme and cobalamin biosynthesis. A P. aeruginosa nirE transposon mutant was not complemented by native cobA encoding the SAM-dependent uroporphyrinogen III methyltransferase involved in cobalamin formation. However, bacterial growth of the nirE mutant was observed when cobA was constitutively expressed by a complementing plasmid, underscoring the special requirement of NirE for heme d1 biosynthesis.

Item Type: Article
Uncontrolled Keywords: heme d biosynthesis,precorrin-2,pseudomonas aeruginosa,sam-dependent uroporphyrinogen iii methyltransferase,uroporphyrinogen iii,biochemistry,molecular biology,cell biology ,/dk/atira/pure/subjectarea/asjc/1300/1303
Faculty \ School: Faculty of Science
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Depositing User: LivePure Connector
Date Deposited: 20 Sep 2022 15:31
Last Modified: 03 Oct 2022 07:34
URI: https://ueaeprints.uea.ac.uk/id/eprint/88528
DOI: 10.1111/j.1742-4658.2009.07306.x

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