Dimethyl Sulfoxide → L-Tyrosine

HRID 2195801Notes

Reaction equation

HRID 2195801 reaction equation
C[O:2][C:3]1[CH:8]=[CH:7][C:6](/[CH:9]=N/NC2C3C(=CC=CC=3)C=NN=2)=[CH:5][CH:4]=1.[NH2:22][C@H:23]([C:31]([OH:33])=[O:32])CC1C=CC=CC=1.CS(C)=O.C(S)[C@@H](O)[C@H](O)CS>C1N(CCO)CCN(CCS(O)(=O)=O)C1>[NH2:22][C@H:23]([C:31]([OH:33])=[O:32])[CH2:9][C:6]1[CH:5]=[CH:4][C:3]([OH:2])=[CH:8][CH:7]=1

Participants

6

Reactant 4

HCID 679Dimethyl SulfoxideHCID 6140L-Phenylalanine7×HCID 446094D-1,4-DithiothreitolHCID 9576791hydralazine 4-anisaldehyde hydrazone

Product 1

HCID 6057L-Tyrosine

Reagents, catalysts & solvents 1

HCID 238314-(2-Hydroxyethyl)-1-piperazineethanesulfonic acidSolvent

Procedure

PAH activity was measured at 25° C. for 1 min essentially as described (BjØrgo E, et al., Eur. J. Biochem. 1988; 257(1):1-10). Tetrameric hPAH (1-2 μg) was incubated in 20 mM Hepes 0.2 M NaCl pH 7.0, containing catalase (0.04 mg/ml), 0.1-1 mM L-Phe, DMSO 2.5% and 0-100 μM of each compound. After 4 min incubation at 25° C., ferrous ammonium sulphate (100 μM) was added, and the reaction was triggered after 1 min by adding BH4 (10-200 μM) in the presence of 5 mM DTT. In some experiments, the preincubation step with L-Phe was omitted and L-Phe was added together with BH4 to analyze the L-Phe induced activation of the enzyme. To determine the kinetic parameters for BH4 and L-Phe, BH4 was used at 0-200 μM (at fixed 1 mM L-Phe) and L-Phe at 0-1 mM (BH4 was fixed at 75 μM). L-Tyr formed was quantified by HPLC and fluorimetric detection (DØskeland A P, et al., J. Biol. Chem. 1984; 259:11242-11248). The saturation curves were fitted to hyperbolic (for BH4) or sigmoidal (for L-Phe) kinetic models by using Sigma Plot v.9.0 (SPSS Inc).

Workup

  1. waitAfter 4 min incubation at 25° C.
  2. waitthe reaction was triggered after 1 min

Sources & evidence

ORD record
ord-33c02def21dc45219cedfdc6f19cb0d7
Source dataset
uspto-grants-2012_12
Patent number
US08338423B2

Reaction notes

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