5'-Adp → Adenosine Monophosphate

HRID 1941735Notes

Reaction equation

HRID 1941735 reaction equation
C(S)CS([O-])(=O)=O.[Na+].[P:9]([O:17][CH2:18][C@H:19]1[O:23][C@@H:22]([N:24]2[C:33]3[N:32]=[CH:31][N:30]=[C:28]([NH2:29])[C:27]=3[N:26]=[CH:25]2)[C@H:21]([OH:34])[C@@H:20]1[OH:35])([O:12]P(O)(O)=O)(=[O:11])[OH:10].C(N(CC(O)=O)CC(O)=O)CN(CC(O)=O)CC(O)=O.C1S/C(=C2/N=C3C(S/2)=CC(=O)C=C3)/N[C@H]1C(O)=O>C(O)C(N)(CO)CO>[CH:31]1[N:30]=[C:28]([NH2:29])[C:27]2[N:26]=[CH:25][N:24]([C@@H:22]3[O:23][C@H:19]([CH2:18][O:17][P:9]([OH:12])([OH:11])=[O:10])[C@@H:20]([OH:35])[C@H:21]3[OH:34])[C:33]=2[N:32]=1

Participants

6

Reactant 4

HCID 60225'-AdpHCID 6049Ethylenediaminetetraacetic AcidHCID 23662354MesnaHCID 178515745HCID 178515745, compound

Product 1

HCID 6083Adenosine Monophosphate

Reagents, catalysts & solvents 1

HCID 6503Tris(hydroxymethyl)aminomethaneSolvent

Procedure

Standard curves for the AK enzyme was prepared as follows. Serial dilutions of purified Sulfolobus acidocaldarius AK from 10 microgrammes/ml to 1 fg/ml were prepared in 50 mM Tris, 25 mM MESNA, pH 7.3. 100 microlitres of enzyme was added to each well of a microtitre plate and 100 microlitres of 135 micromolar ADP 15 mM MgAc, 1 mM EDTA added to each well. Three separate standard curves were prepared for incubation of the assay plate at 30 degrees C., 50 degrees C. and 70 degrees C. for 20 minutes. Following incubation 30 microlitres of luciferin/luciferase reagent (Biothema) was added and the signal read in a luminometer (Orion, Berthold) and the results shown in FIG. 3.

Workup

  1. customStandard curves for the AK enzyme was prepared
  2. additionadded to each well
  3. customThree separate standard curves
  4. customwere prepared for incubation of the assay plate at 30 degrees C
  5. custom50 degrees C
  6. custom70 degrees C
  7. additionwas added

Sources & evidence

ORD record
ord-d11127dce50a41d89a5cb6d853e160cd
Source dataset
uspto-grants-2013_03
Patent number
US08389208B2

Reaction notes

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