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Literature summary for 2.7.8.43 extracted from
- The structure of the neisserial lipooligosaccharide phosphoethanolamine transferase A (LptA) required for resistance to polymyxin (2013), J. Mol. Biol., 425, 3389-3402.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene lptA, recombinant expression of wild-type and soluble (membrane-deleted) C-terminally His6-tagged enzyme in Escherichia coli DELTAdsbA mutant strain JCB571 from plasmids pCMK255 and pCMK527, expression of wild-type enzyme confers resistance to polymyxin in Escherichia coli, while expression of the truncated soluble mutant does not | Neisseria meningitidis |
| recombinant expression of wild-type and selenomethionine-labeled His6-tagged soluble periplasmic domain of the enzyme in Escherichia coli strains CKEC272 and CKEC580 | Neisseria meningitidis |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| purified recombinant His6-tagged soluble catalytic domain of LptA, free and selenomethionine-labeled, or in complex with Zn2+ and a truncated form of substrate 1,2-dihexanoyl-sn-glycero-3-phosphoethanolamine, X-ray diffraction structure determination and analysis at 1.43-1.78 A resolution, molecular replacement. and modeling | Neisseria meningitidis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | generation of a soluble enzyme mutant by deletion of the membrane binding structure. The mutant does not confer resistance to polymyxin in transformed Escherichia coli cells, in contrast to the wild-type LptA | Neisseria meningitidis |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| EDTA | - |
Neisseria meningitidis |  |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | metal ions present in the active site are coordinated to Thr280 and to residues conserved among the family of transferases | Neisseria meningitidis | |
| Mg2+ | required, two Mg2+ ions per enzyme molecule, binding structure model | Neisseria meningitidis | |
| additional information | three bound metal ions (two Zn2+ and a Mg2+) are implicated in catalysis | Neisseria meningitidis | |
| Zn2+ | metal ions present in the active site are coordinated to Thr280 and to residues conserved among the family of transferases, interaction of the Zn2+ with the active site, binding site structure, overview. The second Zn2+ ion is coordinated by the side chains of His383, His465, and one of the oxygen atoms of the phosphate group attached to Thr280. In LptA, two types of coordination around the Zn1 are observed depending on the state of Thr280. In the case of non-phosphorylated Thr280, the coordination about Zn1 is tetrahedral with the fourth ligand being the free phosphate group. In contrast, when Thr280 is phosphorylated, a penta-coordinated metal is observed with the hydroxyl oxygen atom of Thr280 and one of the phosphate oxygen atoms each interacting weakly with the metal | Neisseria meningitidis | |
| Zn2+ | required, one Zn2+ ion per enzyme molecule coordinated tetrahedrally by the side chains of Glu240, Asp452, and His453 and the hydroxyl oxygen atom of Thr280, very strong binding to the enzyme, binding structure model | Neisseria meningitidis | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine + lipid A | Neisseria meningitidis | addition of phosphoethanolamine to the phosphate group at the 1-position of lipid A | 1,2-dipalmitoyl-sn-glycerol + lipid A 1-(2-aminoethyl diphosphate) | - |
? | |
| 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine + lipid A | Neisseria meningitidis | addition of phosphoethanolamine to the phosphate group at the 4'-position of lipid A | 1,2-dipalmitoyl-sn-glycerol + lipid A 4'-(2-aminoethyl diphosphate) | - |
? | |
| diacylphosphatidylethanolamine + lipid A | Neisseria meningitidis | - |
diacylglycerol + lipid A 4'-(2-aminoethyl diphosphate) | - |
? | |
| diacylphosphatidylethanolamine + lipid A | Neisseria meningitidis NMB | - |
diacylglycerol + lipid A 4'-(2-aminoethyl diphosphate) | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Neisseria meningitidis | Q7DD94 | - |
- |
| Neisseria meningitidis | Q7DDQ9 | - |
- |
| Neisseria meningitidis NMB | Q7DDQ9 | - |
- |
Posttranslational Modification
| Posttranslational Modification | Comment | Organism |
|---|---|---|
| phosphoprotein | phosphorylation of the putative nucleophile, Thr280 | Neisseria meningitidis |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant wild-type and selenomethionine-labeled His6-tagged soluble periplasmic domain of the enzyme from Escherichia coli by nickel affinity chromatography | Neisseria meningitidis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 1,2-dihexanoyl-sn-glycero-3-phosphoethanolamine + lipid A | low activity, addition of phosphoethanolamine to the phosphate group at the 1-position of lipid A | Neisseria meningitidis | 1,2-dihexanoyl-sn-glycerol + lipid A 1-(2-aminoethyl diphosphate) | - |
? | |
| 1,2-dihexanoyl-sn-glycero-3-phosphoethanolamine + lipid A | low activity, addition of phosphoethanolamine to the phosphate group at the 4'-position of lipid A | Neisseria meningitidis | 1,2-dihexanoyl-sn-glycerol + lipid A 4'-(2-aminoethyl diphosphate) | - |
? | |
| 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine + lipid A | addition of phosphoethanolamine to the phosphate group at the 1-position of lipid A | Neisseria meningitidis | 1,2-dipalmitoyl-sn-glycerol + lipid A 1-(2-aminoethyl diphosphate) | - |
? | |
| 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine + lipid A | addition of phosphoethanolamine to the phosphate group at the 4'-position of lipid A | Neisseria meningitidis | 1,2-dipalmitoyl-sn-glycerol + lipid A 4'-(2-aminoethyl diphosphate) | - |
? | |
| diacylphosphatidylethanolamine + lipid A | - |
Neisseria meningitidis | diacylglycerol + lipid A 4'-(2-aminoethyl diphosphate) | - |
? | |
| diacylphosphatidylethanolamine + lipid A | - |
Neisseria meningitidis NMB | diacylglycerol + lipid A 4'-(2-aminoethyl diphosphate) | - |
? | |
| additional information | in Neisseria meningitidis, phosphatidylethanolamine typically has acyl chains of C12 and C14 with the first position being occupied with a saturated chain and the second being unsaturated. The recombinant soluble periplasmic domain of the enzyme is active in an aqueous assay but unable to add phoshoethanolamine to lipid A in Escherichia coli strains, lipid A profiles, overview | Neisseria meningitidis | ? | - |
? | |
| additional information | enzyme activity assays on the membrane-deleted LptA are performed using 4-nitrophenyl phosphoethanolamine, p-NPPE, as the substrate analogue, the enzyme is capable of cleaving the phosphoethanolamine portion from the p-NPPE chromogenic substrate | Neisseria meningitidis | ? | - |
? | |
| additional information | enzyme activity assays on the membrane-deleted LptA are performed using 4-nitrophenyl phosphoethanolamine, p-NPPE, as the substrate analogue, the enzyme is capable of cleaving the phosphoethanolamine portion from the p-NPPE chromogenic substrate | Neisseria meningitidis NMB | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| More | secondary-structure ribbon representation of the soluble domain of LptA, and modeling of the three-dimensional structure of LptA, overview. Possible formation of five intramolecular disulfide bonds: Cys276-Cys286, Cys327-Cys331, Cys348-Cys353, Cys402-Cys410, and Cys499-Cys540 | Neisseria meningitidis |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| lipid A PEA transferase | - |
Neisseria meningitidis |
| lipooligosaccharide phosphoethanolamine transferase A | - |
Neisseria meningitidis |
| LptA | - |
Neisseria meningitidis |
| phosphoethanolamine transferase A | - |
Neisseria meningitidis |
| sNMLptA | - |
Neisseria meningitidis |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | ethanolamine transferases are members of the YhjW/YjdB/YijP superfamily | Neisseria meningitidis |
| additional information | analysis of the three-dimensional structure of the soluble catalytic domain of LptA, active-site residues, structure homology, overview | Neisseria meningitidis |
| additional information | homology structure modeling of the enzyme, active-site residues and metal binding structures, crystal structure analysis, overview | Neisseria meningitidis |
| physiological function | the addition of PEA to lipid A by lipid A PEA transferase, LptA, is a major mechanism for resistance to polymyxin in Neisseria meningitidis since this species does not synthesize 4-aminoarabinose. The neisserial lipooligosaccharide phosphoethanolamine transferase A is required for resistance to polymyxin | Neisseria meningitidis |
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