Reaction #939015

ord-1cfbfd07b15548a49aa5a01e196e25f6

Reaction equation

CCCCC/C=C\C\C=C/C=C/C=C/[C@@H](SC[C@H](NC(=O)CC[C@H](N)C(=O)O)C(=O)NCC(=O)O)[C@@H](O)CCCC(=O)O
LTC4
CCCCC/C=C\C\C=C/C=C/C=C/[C@@H]1O[C@H]1CCCC(=O)OC
LTA4 methyl ester
N[C@@H](COB(O)O)C(=O)O
serine borate
→
CCCCC/C=C\C\C=C/C=C/C=C/[C@@H](SC[C@@H](NC(=O)CC[C@H](N)C(=O)O)C(=O)NCC(=O)O)[C@@H](O)CCCC(=O)O
Leukotriene C4

Reactants

CCCCC/C=C\C\C=C/C=C/C=C/[C@@H](SC[C@H](NC(=O)CC[C@H](N)C(=O)O)C(=O)NCC(=O)O)[C@@H](O)CCCC(=O)O
LTC4
CCCCC/C=C\C\C=C/C=C/C=C/[C@@H]1O[C@H]1CCCC(=O)OC
LTA4 methyl ester
N[C@@H](COB(O)O)C(=O)O
serine borate

Reagents

None

Solvents

O=P([O-])([O-])[O-]
phosphate

Conditions

Detailed conditions
See reaction.notes.procedure_details.

Workup

  1. 1
    Otherfor 30 minutes
  2. 2
    Otherat 37° C
  3. 3
    OtherThe reaction was terminated by addition of ice-cold methanol

Procedure

LTC4 synthase prepared as a crude fraction from guinea pig lung was used. The test compounds7were tested in duplicate at a concentration of 300 μg/ml. The test compound and/or vehicle was incubated with 12 μg enzyme, 0.3 μg LTA4 methyl ester, 0.2% (w/w) albumin (to stabilize the product) and 4.5 mM serine borate (to prevent conversion of LTC4 to LTD4) in phosphate buffer pH 7.8 for 30 minutes at 37° C. The reaction was terminated by addition of ice-cold methanol. Formation of LTC4 was quantitated by RIA (radioimmunoassay).

Source

DOI: 10.6084/m9.figshare.5104873.v1Patent: US07252845B2uspto-grants-2007_08