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Compensatory thio–redox interactions between DsbA, CcdA and CcmG unveil the apocytochrome c holdase role of CcmG during cytochrome c maturation
Author(s) -
Turkarslan Serdar,
Sanders Carsten,
Ekici Seda,
Daldal Fevzi
Publication year - 2008
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.2008.06441.x
Subject(s) - dsba , biology , biochemistry , cytochrome , heme , thio , cytochrome c , stereochemistry , chemistry , periplasmic space , enzyme , gene , mitochondrion , escherichia coli
Summary During cytochrome c maturation (Ccm), the DsbA‐dependent thio‐oxidative protein‐folding pathway is thought to introduce a disulphide bond into the haem‐binding motif of apocytochromes c. This disulphide bond is believed to be reduced through a thio‐reductive pathway involving the Ccm components CcdA (DsbD), CcmG and CcmH. Here, we show in Rhodobacter capsulatus that in the absence of DsbA cytochrome c levels were decreased and CcdA or CcmG or the putative glutathione transporter CydDC was not needed for Ccm. This decrease was not due to overproduction of the periplasmic protease DegP as a secondary effect of DsbA absence. In contrast, CcmH was absolutely necessary regardless of DsbA, indicating that compensatory thio–redox interactions excluded it. Remarkably, the double (DsbA–CcmG) and triple (DsbA–CcmG–CcdA) mutants produced cytochromes c at lower levels than the DsbA‐null mutants, unless they contained a CcmG derivative (CcmG*) lacking its thio‐reductive activity. Purified CcmG* can bind apocytochrome c in vitro , revealing for the first time a thiol‐independent, direct interaction between apocytochrome c and CcmG. Furthermore, elimination of the thio–redox components does not abolish cytochrome c production, restricting the number of Ccm components essential for haem–apocyt c ligation per se during Ccm.