BRENDA - Enzyme Database

Septal localization by membrane targeting sequences and a conserved sequence essential for activity at the COOH-terminus of Bacillus subtilis cardiolipin synthase

Kusaka, J.; Shuto, S.; Imai, Y.; Ishikawa, K.; Saito, T.; Natori, K.; Matsuoka, S.; Hara, H.; Matsumoto, K.; Res. Microbiol. 167, 202-214 (2016)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.7.8.B10
gene clsA, recombinant expression of wild-type and deletion mutant enzymes in Escherichia coli strain JM109
Bacillus subtilis
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.7.8.B10
additional information
cardiolipin synthesis is abolished after deleting the last residue, Leu482, in the C-terminal four amino acid residue sequence, Ser-Pro-Ile-Leu, which is highly conserved among bacterial CL synthases. A series of N-terminal, internal, and C-terminal deletion derivatives of ClsA fused to GFP are constructed using plasmid vectors pSG1729 and pSG1154. Construction of a series of GFP-tagged membrane targeting sequence derivatives, GFP-MTS(s), and fusion proteins formed by the C- and N-termini. Integration of the constructed clsA alleles into the amyE locus of wild-type strain 168 and CL-deficient BSF219 strain. Analysis of expression and subcellular localization of the mutant proteins, immunohistochemic detection, overview. The cardiolipin-deficient mutant cells of Bacillus subtilis strain BFS219 is harboring an allele of a defective derivative of clsA. A fusion of GFP to the extreme COOH-terminus does not affect septal localization, i.e. the role in septal localization played by the amphipathic alpha-helices is not affected by GFP-fusion
Bacillus subtilis
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
2.7.8.B10
membrane
septal membranes. Recombinant GFP fusion proteins of enzyme ClsA with N-terminal and internal deletions retain septal localization. Enzyme mutants with deletions starting from the C-terminus (Leu482) cease to localize to the septum once the deletion passes the Ile residue at 448, indicating that the sequence responsible for septal localization is confined within a short distance from the C-terminus. Enzyme ClsA contains a specific region responsible for the septal membrane localization
Bacillus subtilis
16020
-
2.7.8.B10
additional information
two sequences, Ile436-Leu450 and Leu466-Leu478, individually form an amphipathic alpha-helix. The configuration is a membrane targeting sequence (MTS), MTS2 and MTS1, respectively. Either one has the ability to affect septal localization, and each of these sequences by itself localizes to the septum
Bacillus subtilis
-
-
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.8.B10
a phosphatidylglycerol + a phosphatidylglycerol
Bacillus subtilis
-
a cardiolipin + glycerol
-
-
?
2.7.8.B10
a phosphatidylglycerol + a phosphatidylglycerol
Bacillus subtilis 168
-
a cardiolipin + glycerol
-
-
?
2.7.8.B10
additional information
Bacillus subtilis
the enzyme homologue ClsA uses only phosphatidylglycerol as substrate in the synthesis of cardiolipin
?
-
-
-
2.7.8.B10
additional information
Bacillus subtilis 168
the enzyme homologue ClsA uses only phosphatidylglycerol as substrate in the synthesis of cardiolipin
?
-
-
-
2.7.8.B11
a phosphatidylglycerol + a phosphatidylethanolamine
Escherichia coli
-
a cardiolipin + ethanolamine
-
-
?
2.7.8.B11
additional information
Escherichia coli
the enzyme homologue ClsC1 (YmdC) uses both phosphatidylethanolamine and phoshatidylglycerol as substrates for cardiolipin synthesis with the assistance of YmdB (ClsC2), the product of a neighboring gene. Enzyme homologue ClsB may also use some phospholipid other than phosphatidylglycerol
?
-
-
-
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.7.8.B10
Bacillus subtilis
-
-
-
2.7.8.B10
Bacillus subtilis 168
-
-
-
2.7.8.B11
Escherichia coli
-
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.8.B10
a phosphatidylglycerol + a phosphatidylglycerol
-
739625
Bacillus subtilis
a cardiolipin + glycerol
-
-
-
?
2.7.8.B10
a phosphatidylglycerol + a phosphatidylglycerol
-
739625
Bacillus subtilis 168
a cardiolipin + glycerol
-
-
-
?
2.7.8.B10
additional information
the enzyme homologue ClsA uses only phosphatidylglycerol as substrate in the synthesis of cardiolipin
739625
Bacillus subtilis
?
-
-
-
-
2.7.8.B10
additional information
the enzyme homologue ClsA uses only phosphatidylglycerol as substrate in the synthesis of cardiolipin
739625
Bacillus subtilis 168
?
-
-
-
-
2.7.8.B11
a phosphatidylglycerol + a phosphatidylethanolamine
-
739625
Escherichia coli
a cardiolipin + ethanolamine
-
-
-
?
2.7.8.B11
additional information
the enzyme homologue ClsC1 (YmdC) uses both phosphatidylethanolamine and phoshatidylglycerol as substrates for cardiolipin synthesis with the assistance of YmdB (ClsC2), the product of a neighboring gene. Enzyme homologue ClsB may also use some phospholipid other than phosphatidylglycerol
739625
Escherichia coli
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
2.7.8.B10
More
the predicted domain structure of Bacillus subtilis major CL synthase, ClsA (482 residues), shows two HKD motif segments (H222-N239, H400-N417). Probable catalytic sites, which are conserved in the PLD superfamily of enzymes, are: in the middle of ClsA and at the N-terminus, a pair of transmembrane alpha-helices (I3-F24 and W34-H56)
Bacillus subtilis
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.7.8.B10
gene clsA, recombinant expression of wild-type and deletion mutant enzymes in Escherichia coli strain JM109
Bacillus subtilis
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.7.8.B10
additional information
cardiolipin synthesis is abolished after deleting the last residue, Leu482, in the C-terminal four amino acid residue sequence, Ser-Pro-Ile-Leu, which is highly conserved among bacterial CL synthases. A series of N-terminal, internal, and C-terminal deletion derivatives of ClsA fused to GFP are constructed using plasmid vectors pSG1729 and pSG1154. Construction of a series of GFP-tagged membrane targeting sequence derivatives, GFP-MTS(s), and fusion proteins formed by the C- and N-termini. Integration of the constructed clsA alleles into the amyE locus of wild-type strain 168 and CL-deficient BSF219 strain. Analysis of expression and subcellular localization of the mutant proteins, immunohistochemic detection, overview. The cardiolipin-deficient mutant cells of Bacillus subtilis strain BFS219 is harboring an allele of a defective derivative of clsA. A fusion of GFP to the extreme COOH-terminus does not affect septal localization, i.e. the role in septal localization played by the amphipathic alpha-helices is not affected by GFP-fusion
Bacillus subtilis
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
2.7.8.B10
membrane
septal membranes. Recombinant GFP fusion proteins of enzyme ClsA with N-terminal and internal deletions retain septal localization. Enzyme mutants with deletions starting from the C-terminus (Leu482) cease to localize to the septum once the deletion passes the Ile residue at 448, indicating that the sequence responsible for septal localization is confined within a short distance from the C-terminus. Enzyme ClsA contains a specific region responsible for the septal membrane localization
Bacillus subtilis
16020
-
2.7.8.B10
additional information
two sequences, Ile436-Leu450 and Leu466-Leu478, individually form an amphipathic alpha-helix. The configuration is a membrane targeting sequence (MTS), MTS2 and MTS1, respectively. Either one has the ability to affect septal localization, and each of these sequences by itself localizes to the septum
Bacillus subtilis
-
-
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.7.8.B10
a phosphatidylglycerol + a phosphatidylglycerol
Bacillus subtilis
-
a cardiolipin + glycerol
-
-
?
2.7.8.B10
a phosphatidylglycerol + a phosphatidylglycerol
Bacillus subtilis 168
-
a cardiolipin + glycerol
-
-
?
2.7.8.B10
additional information
Bacillus subtilis
the enzyme homologue ClsA uses only phosphatidylglycerol as substrate in the synthesis of cardiolipin
?
-
-
-
2.7.8.B10
additional information
Bacillus subtilis 168
the enzyme homologue ClsA uses only phosphatidylglycerol as substrate in the synthesis of cardiolipin
?
-
-
-
2.7.8.B11
a phosphatidylglycerol + a phosphatidylethanolamine
Escherichia coli
-
a cardiolipin + ethanolamine
-
-
?
2.7.8.B11
additional information
Escherichia coli
the enzyme homologue ClsC1 (YmdC) uses both phosphatidylethanolamine and phoshatidylglycerol as substrates for cardiolipin synthesis with the assistance of YmdB (ClsC2), the product of a neighboring gene. Enzyme homologue ClsB may also use some phospholipid other than phosphatidylglycerol
?
-
-
-
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.7.8.B10
a phosphatidylglycerol + a phosphatidylglycerol
-
739625
Bacillus subtilis
a cardiolipin + glycerol
-
-
-
?
2.7.8.B10
a phosphatidylglycerol + a phosphatidylglycerol
-
739625
Bacillus subtilis 168
a cardiolipin + glycerol
-
-
-
?
2.7.8.B10
additional information
the enzyme homologue ClsA uses only phosphatidylglycerol as substrate in the synthesis of cardiolipin
739625
Bacillus subtilis
?
-
-
-
-
2.7.8.B10
additional information
the enzyme homologue ClsA uses only phosphatidylglycerol as substrate in the synthesis of cardiolipin
739625
Bacillus subtilis 168
?
-
-
-
-
2.7.8.B11
a phosphatidylglycerol + a phosphatidylethanolamine
-
739625
Escherichia coli
a cardiolipin + ethanolamine
-
-
-
?
2.7.8.B11
additional information
the enzyme homologue ClsC1 (YmdC) uses both phosphatidylethanolamine and phoshatidylglycerol as substrates for cardiolipin synthesis with the assistance of YmdB (ClsC2), the product of a neighboring gene. Enzyme homologue ClsB may also use some phospholipid other than phosphatidylglycerol
739625
Escherichia coli
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.7.8.B10
More
the predicted domain structure of Bacillus subtilis major CL synthase, ClsA (482 residues), shows two HKD motif segments (H222-N239, H400-N417). Probable catalytic sites, which are conserved in the PLD superfamily of enzymes, are: in the middle of ClsA and at the N-terminus, a pair of transmembrane alpha-helices (I3-F24 and W34-H56)
Bacillus subtilis
General Information
EC Number
General Information
Commentary
Organism
2.7.8.B10
evolution
the enzyme belongs to the PLD superfamily of enzymes
Bacillus subtilis
2.7.8.B10
additional information
the fact that the homologues YwiE and YwjE show almost no detectable cardiolipin synthesis activity in Bacillus subtilis cells under laboratory conditions, although both localize septally, suggests a much stricter consensus motif S-P-(I/L)-L for bacterial CL synthase, that is, the two residues in the middle of the four conserved residues may have to be taken into account for activity
Bacillus subtilis
General Information (protein specific)
EC Number
General Information
Commentary
Organism
2.7.8.B10
evolution
the enzyme belongs to the PLD superfamily of enzymes
Bacillus subtilis
2.7.8.B10
additional information
the fact that the homologues YwiE and YwjE show almost no detectable cardiolipin synthesis activity in Bacillus subtilis cells under laboratory conditions, although both localize septally, suggests a much stricter consensus motif S-P-(I/L)-L for bacterial CL synthase, that is, the two residues in the middle of the four conserved residues may have to be taken into account for activity
Bacillus subtilis