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Different residues in periplasmic domains of the CcmC inner membrane protein of Pseudomonas fluorescens ATCC 17400 are critical for cytochrome c biogenesis and pyoverdine‐mediated iron uptake
Author(s) -
Gaballa Ahmed,
Baysse Christine,
Koedam Nico,
Muyldermans Serge,
Cornelis Pierre
Publication year - 1998
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.1365-2958.1998.01085.x
Subject(s) - biology , periplasmic space , pseudomonas fluorescens , pyoverdine , biogenesis , cytochrome , siderophore , azurin , inner membrane , bacterial outer membrane , microbiology and biotechnology , biochemistry , pseudomonas , bacteria , membrane , escherichia coli , genetics , gene , electron transfer , enzyme , chemistry , organic chemistry
The inner membrane protein CcmC (CytA) of Pseudomonas fluorescens ATCC17400, which has homologues in several bacteria and plant mitochondria, is needed for the biogenesis of cytochrome c . A CcmC‐deficient mutant is also compromised in the production and utilization of pyoverdine, the high‐affinity fluorescent siderophore. A topological model for CcmC, based on the analysis of alkaline phosphatase fusions, predicts six membrane‐spanning regions with three periplasmic loops. Site‐directed mutagenesis was used in order to assess the importance of some periplasm‐exposed residues, conserved in all CcmC homologues, for cytochrome c biogenesis, and pyoverdine production/utilization. Despite the conservation of the residues His‐61, Val‐62 and Pro‐63 in the first periplasmic loop, and Leu‐184, His‐185 and Gln‐186 in the third periplasmic loop, their simultaneous replacement with Ala only partially affected cytochrome c biogenesis and pyoverdine production/utilization. Simultaneous replacements of residues Trp‐115 and Gly‐116 in the second periplasmic loop substantially affected pyoverdine production/utilization but not cytochrome c production. An Ala substitution of Asp‐127, in the second periplasmic loop, resulted in decreased production of cytochrome c , slower growth in conditions of anaerobiosis and reduced pyoverdine production. On the other hand, a mutation in Trp‐126, also in the second periplasmic loop, totally suppressed the production of cytochrome c , whereas it had no effect on the production and utilization of pyoverdine. These results show a differential involvement of amino acid residues in periplasmic domains of CcmC in cytochrome c biogenesis and pyoverdine production/utilization.