The cytochrome c maturation components CcmF, CcmH, and CcmI form a membrane-integral multisubunit heme ligation complex

Carsten Sanders, Serdar Turkarslan, Dong Woo Lee, Ozlem Onder, Robert G. Kranz, Fevzi Daldal

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Cytochrome c maturation (Ccm) is a post-translational and post-export protein modification process that involves ten (CcmABCDEFGHI and CcdA or DsbD) components in most Gram-negative bacteria. The absence of any of these components abolishes the ability of cells to form cytochrome c, leading in the case of Rhodobacter capsulatus to the loss of photosynthetic proficiency and respiratory cytochrome oxidase activity. Based on earlier molecular genetic studies, we inferred that R. capsulatus CcmF, CcmH, and CcmI interact with each other to perform heme-apocytochrome c ligation. Here, using functional epitope-tagged derivatives of these components coproduced in appropriate mutant strains, we determined protein-protein interactions between them in detergent-dispersed membranes. Reciprocal affinity purification as well as tandem size exclusion and affinity chromatography analyses provided the first biochemical evidence that CcmF, CcmH, and CcmI associate stably with each other, indicating that these Ccm components form a membrane-integral complex. Under the conditions used, the CcmFHI complex does not contain CcmG, suggesting that the latter thio-reduction component is not always associated with the heme ligation components. The findings are discussed with respect to defining the obligatory components of a minimalistic heme-apocytochrome c ligation complex in R. capsulatus.

Original languageEnglish
Pages (from-to)29715-29722
Number of pages8
JournalJournal of Biological Chemistry
Volume283
Issue number44
DOIs
Publication statusPublished - 2008 Oct 31

Fingerprint

Rhodobacter capsulatus
Cytochromes c
Heme
Ligation
Membranes
Affinity chromatography
Proteins
Size exclusion chromatography
Electron Transport Complex IV
Post Translational Protein Processing
Gram-Negative Bacteria
Affinity Chromatography
Detergents
Purification
Gel Chromatography
Epitopes
Molecular Biology
Bacteria
Derivatives

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Sanders, Carsten ; Turkarslan, Serdar ; Lee, Dong Woo ; Onder, Ozlem ; Kranz, Robert G. ; Daldal, Fevzi. / The cytochrome c maturation components CcmF, CcmH, and CcmI form a membrane-integral multisubunit heme ligation complex. In: Journal of Biological Chemistry. 2008 ; Vol. 283, No. 44. pp. 29715-29722.
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The cytochrome c maturation components CcmF, CcmH, and CcmI form a membrane-integral multisubunit heme ligation complex. / Sanders, Carsten; Turkarslan, Serdar; Lee, Dong Woo; Onder, Ozlem; Kranz, Robert G.; Daldal, Fevzi.

In: Journal of Biological Chemistry, Vol. 283, No. 44, 31.10.2008, p. 29715-29722.

Research output: Contribution to journalArticle

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