Cloned (Comment) | Organism |
---|---|
cDNA coding for the predicted mature PLA2 protein, amino acids 1-128 without the signal peptide, designed with optimized codons for high level expression in Escherichia coli strain Tuner (DE3) LacI | Oryza sativa |
Crystallization (Comment) | Organism |
---|---|
isoform 2 phospholipase A2, by the sitting drop, vapor-diffusion method, to 2.0 A resolution. The N-terminal half of the chain contains mainly loop structure, including the conserved Ca2+-binding loop, but starts with a short 3(10)-helix and also includes two short anti-parallel beta-strands. The C-terminal half is folded into three anti-parallel alpha-helices, of which the two first contain the conserved catalytic histidine and calcium liganding aspartate residues. The structure is stabilized by six disulfide bonds. The water structure around the calcium ion binding site suggests the involvement of a second water molecule in the mechanism for hydrolysis, the water-assisted calcium-coordinate oxyanion mechanism. The octanoate molecule in the complex structure is bound in a hydrophobic pocket, which extends to the likely membrane interface and is proposed to model the binding of the product fatty acid | Oryza sativa |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construct A (Leu-1 mutated to an alanine) and construct B (extra glycine inserted between the start methionine and Leu-1), both variants show similar specific activity after refolding | Oryza sativa |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | Ca2+-binding loop | Oryza sativa |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Oryza sativa | Q9XG81 | - |
- |
Purification (Comment) | Organism |
---|---|
by centrifugation, sonication and gel filtration | Oryza sativa |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
33 | - |
construct A (Leu-1 mutated to an alanine) and construct B (extra glycine inserted between the start methionine and Leu-1) | Oryza sativa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-palmitoyl-2-oleoylglycerophosphocholine + H2O | - |
Oryza sativa | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
class XIB phospholipase A2 | - |
Oryza sativa |
sPLA2 | - |
Oryza sativa |