Cloned (Comment) | Organism |
---|---|
pgpB gene is cloned into the pET22b vector (Novagen) to generate the C-terminal His6-tagged protein. Mutants were generated using standard site-directed mutagenesis. The PgpB wild type and mutant proteins are expressed in Escherichia coli strain C41(DE3) | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
vapor diffusion method, the structure is determined at 3.2 A resolution | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D211A | mutant enzyme retains 50% activity | Escherichia coli |
H163A | mutation increases hydrolysis of 1,2-dipalmitoryl-sn-glycero-3-[phospho-rac-(1'-(3'-phospho)glycerol)] by 50% | Escherichia coli |
K93A | the mutant enzyme shows complete activity loss for lysophosphatidic acid and (2S,3R,4E)-2-aminooctadec-4-ene-1,3-diol-1-phosphate, whereas it retains full activity for phosphatidic acid and gains 40% more activity toward phosphatidylglycerol phosphate compared with wild type enzyme | Escherichia coli |
K93A/K97A | the mutant enzyme shows complete activity loss for lysophosphatidic acid and (2S,3R,4E)-2-aminooctadec-4-ene-1,3-diol-1-phosphate, whereas it retains full activity for phosphatidic acid and gained 40% more activity toward phosphatidylglycerol phosphate compared with wild type enzyme | Escherichia coli |
K93T | mutant enzyme with significantly improved activity for lysophosphatidic acid, (2S,3R,4E)-2-aminooctadec-4-ene-1,3-diol-1-phosphate, and phosphatidic acid by 2.5- or 3.4-fold but not for 1,2-dipalmitoryl-sn-glycero-3-[phospho-rac-(1'-(3'-phospho)glycerol)] | Escherichia coli |
K93T/K97A | the mutant enzyme loses all activity for lysophosphatidic acid or (2S,3R,4E)-2-aminooctadec-4-ene-1,3-diol-1-phosphate, but retains 50% activity for phosphatidic acid or 1,2-dipalmitoryl-sn-glycero-3-[phospho-rac-(1'-(3'-phospho)glycerol)] | Escherichia coli |
K97A | the mutant enzyme shows complete activity loss for lysophosphatidic acid and (2S,3R,4E)-2-aminooctadec-4-ene-1,3-diol-1-phosphate, whereas it retains full activity for phosphatidic acid and gained 40% more activity toward phosphatidylglycerol phosphate compared with wild type enzyme | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
orthovanadate | - |
Escherichia coli | |
phosphatidylethanolamine | 3 mM, 70% inhibition. competitive | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.108 | - |
1-(9Z-octadecenoyl)-sn-glycero-3-phosphate | pH 7.5, 23°C | Escherichia coli | |
5.052 | - |
1-hexanoyl-sn-glycero-3-phosphate | pH 7.5, 23°C | Escherichia coli |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | integral membrane protein | Escherichia coli | 16020 | - |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A924 | - |
- |
Escherichia coli K12 | P0A924 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2S,3R,4E)-2-aminooctadec-4-ene-1,3-diol-1-phosphate + H2O | - |
Escherichia coli | ? | - |
? | |
(2S,3R,4E)-2-aminooctadec-4-ene-1,3-diol-1-phosphate + H2O | - |
Escherichia coli K12 | ? | - |
? | |
1,2-dihexadecanoyl-sn-glycero-3-phosphate + H2O | - |
Escherichia coli | 1,2-dihexadecanoyl-sn-glycerol + phosphate | - |
? | |
1,2-dihexadecanoyl-sn-glycero-3-phosphate + H2O | - |
Escherichia coli K12 | 1,2-dihexadecanoyl-sn-glycerol + phosphate | - |
? | |
1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1'-(3'-phospho)glycerol)] + H2O | - |
Escherichia coli | ? + phosphate | - |
? | |
1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1'-(3'-phospho)glycerol)] + H2O | - |
Escherichia coli K12 | ? + phosphate | - |
? | |
1-(9Z-octadecenoyl)-sn-glycero-3-phosphate + H2O | - |
Escherichia coli | 1-(9Z-octadecenoyl)-sn-glycerol + phosphate | - |
? | |
1-(9Z-octadecenoyl)-sn-glycero-3-phosphate + H2O | - |
Escherichia coli K12 | 1-(9Z-octadecenoyl)-sn-glycerol + phosphate | - |
? | |
1-hexanoyl-sn-glycero-3-phosphate + H2O | - |
Escherichia coli | 1-hexanoyl-sn-glycerol + phosphate | - |
? | |
1-hexanoyl-sn-glycero-3-phosphate + H2O | - |
Escherichia coli K12 | 1-hexanoyl-sn-glycerol + phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
PgpB | - |
Escherichia coli |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
2.7 | - |
phosphatidylethanolamine | pH 7.5, 23°C | Escherichia coli |