Expression of the Solfolobus acidocaldarius Rieske iron sulfur protein II (SOXF) with the correctly inserted [2FE-2S] cluster in Escherichia coli

Biochemical and Biophysical Research Communications
C L SchmidtG Schäfer

Abstract

The Rieske protein II (Schmidt et al., 1996, FEBS Lett. 388, 43-46) from the thermoacidophilic crenarcheon Sulfolobus acidocaldarius (DSM 639) was expressed in E. coli cells. The full length protein was strictly bound to the E. coli membranes and could only be removed by detergent treatment indicating the presence of a membrane anchor. The iron sulfur cluster was correctly inserted into a fraction of the full length protein and much more effectively into a soluble form created by the deletion of the 45 N-terminal amino acids. The soluble form of the protein displayed the typical spectroscopic properties of a respiratory Rieske protein. The midpoint potential was +375 mV determined by CD redox potentiometry. The presented data demonstrate that the structure of the recombinant protein is very similar or identical to the authentic protein making this a powerful model system for the studies of Rieske proteins by site directed mutagenesis.

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Citations

Aug 23, 2007·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Heiko BönischRudolf Ladenstein
Feb 21, 2008·Biometals : an International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine·S BoxhammerChristian L Schmidt
Aug 23, 2008·Journal of Biological Inorganic Chemistry : JBIC : a Publication of the Society of Biological Inorganic Chemistry·Eric N BrownS Ramaswamy
Nov 1, 2011·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Hauke PaulsenVolker Schünemann
Jan 15, 2002·The Journal of Biological Chemistry·Dirk SchneiderMatthias Rögner
Jul 13, 2000·Journal of Molecular Biology·M SchützW Nitschke
Jun 11, 2002·Journal of Molecular Biology·Heiko BönischRudolf Ladenstein
May 6, 2003·Protein Expression and Purification·Beata GubernatorAndrzej Szczepaniak
Sep 8, 1999·Microbiology and Molecular Biology Reviews : MMBR·G SchäferV Müller
Feb 7, 2006·Biochimica Et Biophysica Acta·Beata GubernatorAndrzej Szczepaniak

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