Potassium Chloride → HCID 501926, compound

HRID 1164780Notes

Reaction equation

HRID 1164780 reaction equation
N[C@H](C(O)=O)CC1N=CNC=1.[Cl-].[K+].[Na+].[Cl-].[Mg+2].[Cl-].[Cl-].P(OC[C@H]1O[C@@H](N2C3N=CN=C(N)C=3N=C2)[C@H](O)[C@@H]1O)(OP(OP(O)(O)=O)(O)=O)(=O)O.C[C@@H]1O[C@@H]([O:57][C@@H:58]2[CH2:67][C@:66]3(O)[C@@:61](CO)([C@H:62]4[C@H:75](O)[CH2:74][C@@:73]5(C)[C@:69](O)([CH2:70][CH2:71][C@@H:72]5[C:78]5C[O:82][C:80](=[O:81])[CH:79]=5)[C@@H:63]4[CH2:64][CH2:65]3)[C@H:60](O)[CH2:59]2)[C@H](O)[C@H](O)[C@H]1O>O>[CH3:71][CH2:70][CH2:69][CH2:63][CH2:64][CH:65]=[CH:66][CH2:67][C@@H:58]([OH:57])[CH:59]=[CH:60][CH:61]=[CH:62][CH2:75][CH:74]=[CH:73][CH2:72][CH2:78][CH2:79][C:80]([OH:82])=[O:81]

Participants

8

Reactant 6

HCID 4873Potassium ChlorideHCID 5234Sodium ChlorideHCID 59575'-AtpHCID 439501OuabainHCID 5360315Magnesium ChlorideHCID 178419078HCID 178419078, compound

Product 1

HCID 501926HCID 501926, compound

Reagents, catalysts & solvents 1

HCID 962WaterSolvent

Procedure

The activity of Na+ -K+ -ATPase was measured as the rate of release of inorganic phosphate in the presence of 30 mM histidine, 20 mM KCl, 30 mM NaCl, 3 mM MgCl2 and 3 mM ATP, pH 7.5 [Jorgenson, supra]. After equilibration for 5 min. at 37° C., 10 μl of the enzyme preparation (1-5 μg protein) was added to 0.5 ml of reaction mixture. The reaction was allowed to proceed for 30 min. at 37° C. The compounds to be tested were dissolved in PBS buffer and added to the enzyme 10 min. prior to the addition of the reaction mixture. Ouabain was dissolved in distilled water. The reaction was terminated by the addition of 0.8 ml color reagent containing ammonium molybdate malachite green and sterox as described by O. A. Candia et al., Biochim. Biophys. Acta 602, pp. 389-400 (1980). After 1 min., 100 μl of 34% sodium citrate solution was added and mixed. The final solution was then read at 660 mm in a Beckman DB spectrophotometer. Al solutions were read against references containing the same concentration of ATP as the incubation tubes. Enzyme activity was expressed as μmol Pi released/h/mg protein.

Workup

  1. custom10 μl of the enzyme preparation (1-5 μg protein)
  2. additionwas added to 0.5 ml of reaction mixture
  3. waitto proceed for 30 min. at 37° C
  4. dissolutionThe compounds to be tested were dissolved in PBS buffer
  5. additionadded to the enzyme 10 min.
  6. additionto the addition of the reaction mixture
  7. customThe reaction was terminated by the addition of 0.8 ml color reagent
  8. additioncontaining ammonium molybdate malachite green and sterox
  9. waitAfter 1 min.
  10. addition100 μl of 34% sodium citrate solution was added
  11. additionmixed
  12. additioncontaining
  13. concentrationthe same concentration of ATP as the incubation tubes

Sources & evidence

ORD record
ord-be901c5c2e4548129b01d94de778a81a
Source dataset
uspto-grants-1990_03
Patent number
US04906467

Reaction notes

Loading…