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Paracoccus denitrificans CcmG is a periplasmic protein–disulphide oxidoreductase required for c ‐ and aa 3 ‐type cytochrome biogenesis; evidence for a reductase role in vivo
Author(s) -
Page M. Dudley,
Ferguson Stuart J.
Publication year - 1997
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 - 10.1046/j.1365-2958.1997.4061775.x
Subject(s) - paracoccus denitrificans , biology , biochemistry , periplasmic space , cytochrome c , cytochrome c oxidase , cytochrome , oxidoreductase , electron transport complex iv , coenzyme q – cytochrome c reductase , microbiology and biotechnology , escherichia coli , enzyme , mitochondrion , gene
Cloning and sequencing of the Paracoccus denitrificans ccmG gene indicates that it codes for a periplasmic protein–disulphide oxidoreductase; the presence of the sequence Cys‐Pro‐Pro‐Cys at the CcmG active site suggests that it may act in vivo to reduce disulphide bonds rather than to form them. A CcmG–PhoA fusion confirmed the periplasmic location. Disruption of the ccmG gene resulted in not only the expected phenotype of pleiotropic deficiency in c ‐type cytochromes, but also loss of spectroscopically detectable cytochrome aa 3 , cytochrome c oxidase and ascorbate/TMPD oxidase activities; there was also an enhanced sensitivity to growth inhibition by some component of rich media and by oxidized thiol compounds. Dithiothreitol promoted the growth of the ccmG mutant on rich media and substantially restored spectroscopically detectable cytochrome aa 3 and cytochrome c oxidase activity, although it did not restore c ‐type cytochrome biogenesis. Assembly of the disulphide‐bridged proteins methanol dehydrogenase and Escherichia coli alkaline phosphatase was unaffected in the ccmG mutant. It is proposed that P. denitrificans CcmG acts in vivo to reduce protein–disulphide bonds in certain protein substrates including c ‐type cytochrome polypeptides and/or polypeptides involved in c ‐type cytochrome biogenesis.