Dimethyl Sulfoxide → Prostaglandin E2

HRID 2118007

Reaction equation

HRID 2118007 reaction equation
CS(C)=O.C(N(CC(O)=O)CC(O)=O)CN(CC(O)=O)CC(O)=O.C([C@H](N)C(O)=O)CC(N[C@H](C(NCC(O)=O)=O)CS)=O.[CH3:45][CH2:46][CH2:47][CH2:48][CH2:49][C@H:50]([OH:69])/[CH:51]=[CH:52]/[C@@H:53]1[C@@H:57]([CH2:58]/[CH:59]=[CH:60]\[CH2:61][CH2:62][CH2:63][C:64]([OH:66])=[O:65])[C@H:56]2[O:67][O:68][C@@H:54]1[CH2:55]2>C(O)(C)C>[CH3:45][CH2:46][CH2:47][CH2:48][CH2:49][C@H:50]([OH:69])/[CH:51]=[CH:52]/[C@@H:53]1[C@@H:57]([CH2:58]/[CH:59]=[CH:60]\[CH2:61][CH2:62][CH2:63][C:64]([OH:66])=[O:65])[C:56](=[O:67])[CH2:55][C@H:54]1[OH:68]

Participants

6

Reactant 4

HCID 679Dimethyl SulfoxideHCID 6049Ethylenediaminetetraacetic AcidHCID 124886GlutathioneHCID 445049Prostaglandin H2

Product 1

HCID 5280360Prostaglandin E2

Reagents, catalysts & solvents 1

HCID 3776IsopropanolSolvent

Procedure

Prostaglandin E synthase microsomal fractions are prepared from CHO—K1 cells transiently transfected with plasmid encoding the human mPGES-1 cDNA. Microsomes are then prepared and the PGES assay begins with the incubation of 5 μg/ml microsomal PGES-1 with compound or DMSO (final 1%) for 20-30 minutes at room temperature. The enzyme reactions are performed in 200 mM KPi pH 7.0, 2 mM EDTA and 2.5 mM GSH-reduced form. The enzymatic reaction is then initiated by the addition of 1 μM final PGH2 substrate prepared in isopropanol (3.5% final in assay well) and incubated at room temperature for 30 seconds. The reaction is terminated by the addition of SnCl2 in 1N HCl (1 mg/ml final). Measurement of PGE2 production in the enzyme reaction aliquots is done by EIA using a standard commercially available kit (Cat #: 901-001 from Assay Designs).

Workup

  1. customMicrosomes are then prepared
  2. customfor 20-30 minutes
  3. customat room temperature
  4. customThe enzymatic reaction
  5. customThe reaction is terminated by the addition of SnCl2 in 1N HCl (1 mg/ml final)
  6. customMeasurement of PGE2 production in the enzyme reaction aliquots

Sources & evidence

ORD record
ord-20f7d08a4bfb4f4db089fbe36ff3c990
Source dataset
uspto-grants-2008_10
Patent number
US07442716B2

Linked notes

Loading…