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Rhodobacter capsulatus magnesium chelatase subunit BchH contains an oxygen sensitive iron–sulfur cluster

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Abstract

Magnesium chelatase is the first unique enzyme of the bacteriochlorophyll biosynthetic pathway. It consists of three subunits (BchI, BchD, and BchH). Amino acid sequence analysis of the Rhodobacter capsulatus BchH revealed a novel cysteine motif (393CX2CX3CX14C) that was found in only six other proteobacteria (CX2CX3CX11–14C). The cysteine motif is likely to coordinate an unprecedented [Fe–S] cluster. Purified BchH demonstrated absorbance in the 460 nm region. This absorbance was abolished in BchH proteins with alanine substitutions at positions Cys396 and Cys414. These modified proteins were also EPR silent. In contrast, wild type BchH protein in the reduced state showed EPR signals resembling those of a [4Fe–4S] cluster with rhombic symmetry and g values at 1.90, 1.93, and 2.09, superimposed with a [3Fe–4S] cluster centered at g = 2.02. The [3Fe–4S] signal was observed independently of the [4Fe–4S] signal under oxidizing conditions. Mg-chelatase activity assays showed that the cluster is not catalytic. We suggest that the [4Fe–4S] and [3Fe–4S] signals originate from a single coordination site on the monomeric BchH protein and that the [4Fe–4S] cluster is sensitive to oxidation. It is speculated that the cluster participates in the switching between aerobic and anaerobic life of the proteobacteria.

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Abbreviations

AAA:

ATPase associated with various cellular activities

ALA:

Aminolevulinate

DTT:

Dithiothreitol

IPTG:

Isopropyl β-d-1-thiogalactopyranoside

MgPME:

Magnesium protoporphyrin monomethylester

PMS:

Phenazine methosulfate

PMSF:

Phenylmethylsulfonyl fluoride

PnSB:

Purple nonsulfur bacteria

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Acknowledgments

We thank Dr. Cecilia Hagerhäll for fruitful discussions. NS greatly acknowledges a fellowship from the Sven and Lilly Lawski Foundation. This work was supported by the Swedish Research Council (MH, FM, and SS), the Swedish National Energy Administration, DESS and the Knut and Alice Wallenberg Foundation (FM and SS).

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Authors and Affiliations

  1. Department of Biochemistry, Center for Molecular Protein Science, Lund University, P.O. Box 124, 221 00, Lund, Sweden

    Nick Sirijovski, Ulf Olsson & Mats Hansson

  2. Department of Photochemistry and Molecular Science, Uppsala University, Villavägen 6, 752 36, Uppsala, Sweden

    Fikret Mamedov & Stenbjörn Styring

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  1. Nick Sirijovski
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  2. Fikret Mamedov
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Correspondence to Nick Sirijovski.

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Communicated by Theo Hansen.

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Sirijovski, N., Mamedov, F., Olsson, U. et al. Rhodobacter capsulatus magnesium chelatase subunit BchH contains an oxygen sensitive iron–sulfur cluster. Arch Microbiol 188, 599–608 (2007). https://doi.org/10.1007/s00203-007-0282-1

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