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Methods for Nitrogenase-Like Dark Operative Protochlorophyllide Oxidoreductase

  • Jürgen MoserEmail author
  • Markus J. Bröcker
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 766)

Abstract

Nitrogenase-like dark operative protochlorophyllide oxidoreductase (DPOR) is involved in the biosynthesis of chlorophylls and bacteriochlorophylls in gymnosperms, ferns, algae, and photosynthetic bacteria. During protochlorophyllide (Pchlide) reduction, the homodimeric subunit ChlL2 of DPOR transfers electrons on the corresponding heterotetrameric catalytic subunit (ChlN/ChlB)2. Although DPOR shares significant amino acid sequence homology to the nitrogenase system, only the initial catalytic steps of DPOR resemble nitrogenase catalysis. Investigation of the cyanobacterial DPOR from Prochlorococcus marinus indicated that subcomplex ChlL2 is functioning as an ATP-dependent switch protein, triggering the transient interaction of ChlL2 and (ChlN/ChlB)2. This dynamic subunit interplay is responsible for the transfer of a single electron from the [4Fe–4S] cluster of ChlL2 onto a second [4Fe–4S] cluster located on (ChlN/ChlB)2. However, the second part of DPOR catalysis is unrelated to nitrogenase catalysis, since no molybdenum-containing cofactor or a P-cluster equivalent is employed. Instead, two consecutive electron transfer steps are mediated via the [4Fe–4S] cluster of (ChlN/ChlB)2, resulting in the reduction of the conjugated ring system of the substrate molecule Pchlide (Figs. 5.1a and 5.2).

Key words

Dark operative protochlorophyllide oxidoreductase (DPOR) nitrogenase-like enzyme chlorophyll biosynthesis dynamic switch protein 

References

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    Brocker MJ, Waetzlich D, Saggu M et al (2010) Biosynthesis of (bacterio)chlorophylls: ATP-dependent transient subunit interaction and electron transfer of dark operative protochlorophyllide oxidoreductase. J Biol Chem 285:8268–8277PubMedCrossRefGoogle Scholar
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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Institut für Mikrobiologie, Technische Universität BraunschweigBraunschweigGermany

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