Cloned (Comment) | Organism |
---|---|
expression of C-terminally His6-tagged enzyme in Escherichia coli strain Rosetta BL21(DE3) | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified His-tagged recombinant enzyme, in complex with 9,11-epoxymethano prostaglandin H2, hanging drop vapor diffusion method, condition A: 0.1 M potassium thiocyanate and 30% PEG-MME 2000 in 1:1 protein-reservoir ratio, 5 days, 4°C, condition B: 1.4 M tri-sodium citrate, pH 6.5, using a similar method except in 2:1 protein-reservoir ratio. Micro-crystals from condition A are used to seed crystallization of ligand-free L-PGDS in the same condition but in the absence of SA U44069. Cryoprotection of condition A crystals using reservoir with 25% glycerol added, while crystals from condition B are cryo-protected with 1.6 M tri-sodium citrate solution. X-ray diffraction structure analysis at 1.88-2.09 A resolution | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(5Z,13E,15S)-9alpha,11alpha-epidioxy-15-hydroxyprosta-5,13-dienoate | Homo sapiens | - |
(5Z,13E,15S)-9alpha,15-dihydroxy-11-oxoprosta-5,13-dienoate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P41222 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant C-terminally His6-tagged enzyme from Escherichia coli strain Rosetta BL21(DE3) by nickel affinity chromatography and gel filtration | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(5Z,13E,15S)-9alpha,11alpha-epidioxy-15-hydroxyprosta-5,13-dienoate | - |
Homo sapiens | (5Z,13E,15S)-9alpha,15-dihydroxy-11-oxoprosta-5,13-dienoate | - |
? | |
(5Z,13E,15S)-9alpha,11alpha-epidioxy-15-hydroxyprosta-5,13-dienoate | i.e. prostaglandin H2, PGH2 | Homo sapiens | (5Z,13E,15S)-9alpha,15-dihydroxy-11-oxoprosta-5,13-dienoate | i.e. prostaglandin D2, PGD2 | ? | |
9,11-epoxymethano-prostaglandin H2 | substrate analogue U44069 | Homo sapiens | 9,11-epoxymethano-prostaglandin D2 | product analogue 12415 | ? |
Subunits | Comment | Organism |
---|---|---|
More | analysis of enzyme structure in complex with substrate analogue U44069, 9,11-epoxymethano-PGH2, and in ligand-free form, NMR analysis, overview. The enzyme overall structure shows the classical lipocalin fold comprising an eight-stranded beta-barrel with two alpha-helices. The ligand-bound and the ligand-free structures differ only in the Omega loop conformationss | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
L-PGDS | - |
Homo sapiens |
lipocalin prostaglandin D synthase | - |
Homo sapiens |
General Information | Comment | Organism |
---|---|---|
additional information | analysis of enzyme structure in complex with substrate analogue U44069, 9,11-epoxymethano-PGH2, and in ligand-free form. The catalytic Cys 65 thiol group occurs in two different conformations, each making a distinct hydrogen bond network to neighboring residues, mechanism of the cysteine nucleophile activation, and modelling of dynamics of protein-substrate and protein-product interactions, overview | Homo sapiens |
physiological function | lipocalin prostaglandin D synthase regulates synthesis of an important inflammatory and signaling mediator, prostaglandin D2 | Homo sapiens |