Wheat mitochondria ccmB encodes the membrane domain of a putative ABC transporter involved in cytochrome c biogenesis

The CXXCH motif is usually recognized in the bacterial periplasm as a haem attachment site in apocytochromes c. There is evidence that the Escherichia coli Ccm (cytochrome c maturation) system recognizes little more than the CXXCH sequence. A limited number of periplasmic proteins have this motif and yet are not c-type cytochromes. To explore how unwanted haem attachment to CXXCH might be avoided, and to determine whether haem attachment to the surface of a non-cytochrome protein would be possible, we converted the active-site CXXCK motif of a thioredoxin-like protein into CXXCH, the C-terminal domain of the transmembrane oxidoreductase DsbD (cDsbD). The E. coli Ccm system was found to catalyse haem attachment to a very small percentage of the resultant protein (∼0.2%). We argue that cDsbD folds sufficiently rapidly that only a small fraction fails to avoid the Ccm system, in contrast with bona fide c-type cytochromes that only adopt their tertiary structure following haem attachment. We also demonstrate covalent haem attachment at a low level in vivo to the periplasmic disulfide isomerase DsbC, which contains a native CXXCH motif. These observations provide insight into substrate recognition by the Ccm system and expand our understanding of the requirements for covalent haem attachment to proteins. The possible evolutionary relationship between thioredoxins and c-type cytochromes is discussed.

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Characterization of the cycHJKL genes involved in cytochrome c biogenesis and symbiotic nitrogen fixation in Rhizobium leguminosarum.

Journal of Bacteriology ◽

10.1128/jb.177.17.4927-4934.1995 ◽

1995 ◽

Vol 177 (17) ◽

pp. 4927-4934 ◽

Cited By ~ 27

Author(s):

M J Delgado ◽

K H Yeoman ◽

G Wu ◽

C Vargas ◽

A E Davies ◽

...

Keyword(s):

Nitrogen Fixation ◽

Cytochrome C ◽

Rhizobium Leguminosarum ◽

Symbiotic Nitrogen Fixation ◽

Cytochrome C Biogenesis

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New PCR primers targeting hydrazine synthase and cytochrome c biogenesis proteins in anammox bacteria

Applied Microbiology and Biotechnology ◽

10.1007/s00253-016-8013-7 ◽

2016 ◽

Vol 101 (3) ◽

pp. 1267-1287 ◽

Cited By ~ 11

Author(s):

Zhichao Zhou ◽

Jing Chen ◽

Han Meng ◽

Volodymyr Dvornyk ◽

Ji-Dong Gu

Keyword(s):

Cytochrome C ◽

Pcr Primers ◽

Anammox Bacteria ◽

Cytochrome C Biogenesis

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Cytochrome c biogenesis is involved in the transposon Tn5-mediated bleomycin resistance and the associated fitness effect in Escherichia coli

Molecular Microbiology ◽

10.1046/j.1365-2958.1998.00755.x ◽

2002 ◽

Vol 28 (1) ◽

pp. 15-24 ◽

Cited By ~ 13

Author(s):

Eric Adam ◽

Michael R. Volkert ◽

Michel Blot

Keyword(s):

Escherichia Coli ◽

Cytochrome C ◽

Transposon Tn5 ◽

Fitness Effect ◽

Cytochrome C Biogenesis

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In vitro reconstitution reveals major differences between human and bacterial cytochrome c synthases

eLife ◽

10.7554/elife.64891 ◽

2021 ◽

Vol 10 ◽

Author(s):

Molly C Sutherland ◽

Deanna L Mendez ◽

Shalon E Babbitt ◽

Dustin E Tillman ◽

Olga Melnikov ◽

...

Keyword(s):

Cytochrome C ◽

Heme Proteins ◽

Alpha Helix ◽

Release Mechanism ◽

Cytochrome C Biogenesis ◽

Peptide Analogs ◽

Apocytochrome C ◽

In Vitro Reconstitution ◽

Cytochromes C

Cytochromes c are ubiquitous heme proteins in mitochondria and bacteria, all possessing a CXXCH (CysXxxXxxCysHis) motif with covalently attached heme. We describe the first in vitro reconstitution of cytochrome c biogenesis using purified mitochondrial (HCCS) and bacterial (CcsBA) cytochrome c synthases. We employ apocytochrome c and peptide analogs containing CXXCH as substrates, examining recognition determinants, thioether attachment, and subsequent release and folding of cytochrome c. Peptide analogs reveal very different recognition requirements between HCCS and CcsBA. For HCCS, a minimal 16-mer peptide is required, comprised of CXXCH and adjacent alpha helix 1, yet neither thiol is critical for recognition. For bacterial CcsBA, both thiols and histidine are required, but not alpha helix 1. Heme attached peptide analogs are not released from the HCCS active site; thus, folding is important in the release mechanism. Peptide analogs behave as inhibitors of cytochrome c biogenesis, paving the way for targeted control.

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