Cloned (Comment) | Organism |
---|---|
- |
Plexaura homomalla |
Crystallization (Comment) | Organism |
---|---|
deletion mutant of 8R-LOX crystallized by sitting drop vapor diffusion, to 1.85 A resolution, belongs to space group P21 with four molecules in the asymmetric unit. U-shaped channel in 8R-LOX | Plexaura homomalla |
Protein Variants | Comment | Organism |
---|---|---|
I433A | absence of the Ile side chain destabilizes the roof of the U-shaped channel, measurable activity only in the presence of CaCl2 and the detergent emolphogen | Plexaura homomalla |
I433W | has no measurable activity, presumably because the Trp side chain effectively blocks the arachidonic acid binding site | Plexaura homomalla |
L432A | less than 5% of the activity of the wild-type | Plexaura homomalla |
L432F | less than 5% of the activity of the wild-type | Plexaura homomalla |
L432I | less than 5% of the activity of the wild-type | Plexaura homomalla |
L432V | less than 20% of the activity of the wild-type | Plexaura homomalla |
additional information | deletion mutant lacks one of the loops, as well as chelating amino acids from two of the three Ca2+ binding sites (the center site and that most distal from the catalytic domain). The Ca2+ site proximal to the catalytic domain, defined primarily by main chain contacts, remains intact and occupied in the mutant structure. Deletion mutant displays wild-type activity in a membrane-free assay, but Ca2+ does not promote membrane binding of the mutant and does not stimulate enzyme activity in a membrane-based assay | Plexaura homomalla |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.018 | - |
arachidonate | mutant L432I | Plexaura homomalla | |
0.028 | - |
arachidonate | mutant L432V | Plexaura homomalla | |
0.033 | - |
arachidonate | mutant L432A | Plexaura homomalla | |
0.05 | - |
arachidonate | wild-type | Plexaura homomalla | |
0.135 | - |
arachidonate | mutant I433A | Plexaura homomalla |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | has three Ca2+ binding sites flanked by putative membrane insertion loops in the C2-like domain | Plexaura homomalla |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Plexaura homomalla | O16025 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Plexaura homomalla |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
arachidonate + O2 | - |
Plexaura homomalla | (5Z,9E,11Z,14Z)-(8R)-8-hydroperoxyicosa-5,9,11,14-tetraenoate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
8R-lipoxygenase | - |
Plexaura homomalla |
8R-LOX | - |
Plexaura homomalla |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
7 | - |
arachidonate | mutant L432I | Plexaura homomalla | |
12 | - |
arachidonate | mutant L432A | Plexaura homomalla | |
19 | - |
arachidonate | mutant I433A | Plexaura homomalla | |
42 | - |
arachidonate | mutant L432V | Plexaura homomalla | |
206 | - |
arachidonate | wild-type | Plexaura homomalla |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Plexaura homomalla |