A proteomic study of Corynebacterium glutamicum AAA+ protease FtsH.
BACKGROUND
The influence of the membrane-bound AAA+ protease FtsH on membrane and cytoplasmic proteins of Corynebacterium glutamicum was investigated in this study. For the analysis of the membrane fraction, anion exchange chromatography was combined with SDS-PAGE, while the cytoplasmic protein fraction was studied by conventional two-dimensional gel electrophoresis.


RESULTS
In contrast to the situation in other bacteria, deletion of C. glutamicum ftsH has no significant effect on growth in standard minimal medium or response to heat or osmotic stress. On the proteome level, deletion of the ftsH gene resulted in a strong increase of ten cytoplasmic and membrane proteins, namely biotin carboxylase/biotin carboxyl carrier protein (accBC), glyceraldehyde-3-phosphate dehydrogenase (gap), homocysteine methyltransferase (metE), malate synthase (aceB), isocitrate lyase (aceA), a conserved hypothetical protein (NCgl1985), succinate dehydrogenase A (sdhA), succinate dehydrogenase B (sdhB), succinate dehydrogenase CD (sdhCD), and glutamate binding protein (gluB), while 38 cytoplasmic and membrane-associated proteins showed a decreased abundance. The decreasing amount of succinate dehydrogenase A (sdhA) in the cytoplasmic fraction of the ftsH mutant compared to the wild type and its increasing abundance in the membrane fraction indicates that FtsH might be involved in the cleavage of a membrane anchor of this membrane-associated protein and by this changes its localization.


CONCLUSIONS
The data obtained hint to an involvement of C. glutamicum FtsH protease mainly in regulation of energy and carbon metabolism, while the protease is not involved in stress response, as found in other bacteria.
Lüdke
Alja
A
Institut für Biochemie, Universität zu Köln, Zülpicher Str, 47, 50674 Köln, Germany. alja.luedke@uni-koeln.de <alja.luedke@uni-koeln.de>
Krämer
Reinhard
R
Burkovski
Andreas
A
Schluesener
Daniela
D
Poetsch
Ansgar
A
D000596:Amino Acids / Q000096:biosynthesis; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002244:Carbon / Q000378:metabolism; D048230:Corynebacterium glutamicum / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D003593:Cytoplasm / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D004734:Energy Metabolism; D017353:Gene Deletion; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D053858:Metabolic Networks and Pathways; D045726:Metalloproteases / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics
D000596:Amino Acids; D001426:Bacterial Proteins; D008565:Membrane Proteins; D002244:Carbon; D045726:Metalloproteases