Escherichia coli has three cardiolipin synthases. Cardiolipin synthase A encoded by clsA, cardiolipin synthase B encoded by clsB and cardiolipin synthase C, encoded by clsC. Triple deletions of clsA, clsB, and clsC results in the complete depletion of cardiolipin synthase activity in Escherichia coli cells. The double mutant DELTAclsAB mutant still makes cardiolipin in the stationary phase
a minor decrease of cardiolipin content is observed in the ClsA overexpression strain. Phosphatidylethanolamine and phosphatidylglycerol levels remain unaltered upon overexpression of ClsA. ClsA deletion leads to abolishment of phosphytidylcholine-dependent phosphatidylalcohol formation
Escherichia coli has three cardiolipin synthases. Cardiolipin synthase A encoded by clsA, cardiolipin synthase B encoded by clsB and cardiolipin synthase C, encoded by clsC. Triple deletions of clsA, clsB, and clsC results in the complete depletion of cardiolipin synthase activity in Escherichia coli cells. The double mutant DELTAclsAB mutant still makes cardiolipin in the stationary phase
a modest increase of cardiolipin content is observed in the ClsB overexpression strain. Overexpression of ClsB also leads to an increase of phosphatidylethanolamine from 67% to 79% and a decrease of phosphatidylglycerol content from 31% to 14% of phospholipids. Overexpression of ClsB leads to formation of phosphatidylalcohols whereas levels of phosphatidylalcohols are unaltered in the clsB knockout mutant
ClsA and proteins YdhP, YjbJ interact with transporter ProP. All three proteins are concentrated at the cell poles, but only ClsA localization was cardiolipin-dependent. ClsA is N-terminally processed and membrane-anchored, with dual, cytoplasmic, catalytic domains
ClsB additionally catalyzes an alternative mechanism for phosphatidylglycerol synthesis that is PgsA-independent. The reaction in vivo and in vitro converts phosphatidylethanolamine and glycerol into phosphatidylglycerol. When the growth medium is supplemented with glycerol, the expression ClsB significantly increases phosphatidylglycerol and cardiolipin levels, with the growth deficiency of PgsA null strain also being complemented under such conditions
an EYMPE epitope tag is introduced into the interior of Cardiolipin synthase. The tagged polypeptide retains the biological properties of wild type enzyme, including full enzymatic activity. Site-directed mutagenesis is used to alter conserved residues in the N-terminal region. An tagged cardiolipin synthase in which Leu7 and Val8 are both replaced by Ser residues retains in vitro activity but loses most of its in vivo activity. The mutant protein has a higher apparent molecular mass than its parent protein. That conserved residues L7 and V8 play a role in polypeptide processing, topology, or both
cls has about 100 residues at the N-terminus that are missing from the polypeptide specified by the homologous gene f413 (cardiolipin synthase B). The f413 protein catalyzes cardiolipin formation in vitro but not in vivo. Amino acid residues specified by the first 60 codons in cls are not essential for catalytic activity
cls has about 100 residues at the N-terminus that are missing from the polypeptide specified by the homologous gene f413 (cardiolipin synthase B). The f413 protein catalyzes cardiolipin formation in vitro but not in vivo. Amino acid residues specified by the first 60 codons in cls are not essential for catalytic activity
cls has about 100 residues at the N-terminus that are missing from the polypeptide specified by the homologous gene f413 (cardiolipin synthase B). The f413 protein catalyzes cardiolipin formation in vitro but not in vivo. Amino acid residues specified by the first 60 codons in cls are not essential for catalytic activity