Tyrosine detecting

Phenylpyruvic acid can cause mental retardation m infants who are deficient m the enzymes necessary to convert l phenylalanine to l tyrosine This disorder is called phenylketonuria, or PKU disease PKU disease can be detected by a simple test rou tmely administered to newborns It cannot be cured but is controlled by restricting the dietary intake of l phenylalanine In practice this means avoiding foods such as meat that are rich m l phenylalanine... 

This led to the conclusion that these amino acids were essential for the resolution capability and only 6 new libraries of 18 compounds had to be synthesized with these amino acid residues to define the position 3. Surprisingly, the separation abilities of all six libraries were very similar. Therefore, tyrosine was chosen for continuing deconvolution, since it is convenient as its aromatic ring can easily be detected by UV spectrometry. The last step, defining position 5, required the synthesis and testing of 6 individual hexapeptides. 

Figure 4.4 Release of amino acids from cortical slices exposed to 50 mM K+. Measurements by HPEC and fluorescence detection after reaction of amino acids with o-phthalaldehyde 1, aspartate 2, glutamate 3, asparagine 4, serine 5, glutamine 6, histidine 7, homoserine (internal standard) 8, glycine 9, threonine 10, arginine 11, taurine 12, alanine 13, GABA 14, tyrosine. Glutamate concentration is almost 1 pmol/gl which represents a release rate of 30 pmol/min/mg tissue...

By contrast, the cytoplasmic decarboxylation of dopa to dopamine by the enzyme dopa decarboxylase is about 100 times more rapid (Am 4x 10 " M) than its synthesis and indeed it is difficult to detect endogenous dopa in the CNS. This enzyme, which requires pyridoxal phosphate (vitamin B6) as co-factor, can decarboxylate other amino acids (e.g. tryptophan and tyrosine) and in view of its low substrate specificity is known as a general L-aromatic amino-acid decarboxylase. 

Liquid chromatography/electrochemistry (LCEC) has become recognized as a powerful tool for the trace determination of easily oxidizable and reducible compounds. This is because detection of as little as 0.1 pmol of material is readily accomplished with relatively simple and inexpensive equipment. Initial interest in LCEC was generated by the determination of several aromatic matabolites of tyrosine in the central nervous system. However, the application of LCEC into other areas of biochemistry has begun at a growing pace. A bibliography of LCEC applications is available... 

Because LCEC had its initial impact in neurochemical analysis, it is not, surprising that many of the early enzyme-linked electrochemical methods are of neurologically important enzymes. Many of the enzymes involved in catecholamine metabolism have been determined by electrochemical means. Phenylalanine hydroxylase activity has been determined by el trochemicaUy monitoring the conversion of tetrahydro-biopterin to dihydrobiopterin Another monooxygenase, tyrosine hydroxylase, has been determined by detecting the DOPA produced by the enzymatic reaction Formation of DOPA has also been monitored electrochemically to determine the activity of L-aromatic amino acid decarboxylase Other enzymes involved in catecholamine metabolism which have been determined electrochemically include dopamine-p-hydroxylase phenylethanolamine-N-methyltransferase and catechol-O-methyltransferase . Electrochemical detection of DOPA has also been used to determine the activity of y-glutamyltranspeptidase The cytochrome P-450 enzyme system has been studied by observing the conversion of benzene to phenol and subsequently to hydroquinone and catechol... 

Lequea et al. used the activity of tyrosine apodecarboxylase to determine the concentration of the enzyme cofactor pyridoxal 5 -phosphate (vitamin B6). The inactive apoenzyme is converted to the active enzyme by pyridoxal 5 -phosphate. By keeping the cofactor the limiting reagent in the reaction by adding excess apoenzyme and substrate, the enzyme activity is a direct measure of cofactor concentration. The enzymatic reaction was followed by detecting tyramine formation by LCEC. The authors used this method to determine vitamin B6 concentrations in plasma samples. 

Figure 3. Sketch of DDC-expressing neurons in the Drosophila larval CNS. The CNS consists of brain lobes and a segmented ventral ganglion. Filled circles represent dopamine cells open circles represent serotonin cells grayed circles represent DDC cells that contain no detectable tyrosine hydroxylase or serotonin immunoreactivity, indicating that these cells may produce neither transmitter (Lundell and Hirsh, 1994). M, medial dopamine neurons VL, ventrolateral serotonin neurons DL, dorsolateral dopamine neurons. The hatched rectangle shows the region of the ventral ganglion that is shown in Figures 4 and 6.

The synthesized CPMV-alkyne 42 was subjected to the CuAAC reaction with 38. Due to the strong fluorescence of the cycloaddition product 43 as low as 0.5 nM, it could be detected without the interference of starting materials. TMV was initially subjected to an electrophilic substitution reaction at the ortho-position of the phenol ring of tyrosine-139 residues with diazonium salts to insert the alkyne functionality, giving derivative 44 [100]. The sequential CuAAC reaction was achieved with greatest efficiency yielding compound 45, and it was found that the TMV remained intact and stable throughout the reaction. 

H. Mino and T. Ono, Applications of pulsed ELDOR-detected NMR measurements to studies of photosystem II Magnetic characterization of Yd tyrosine radical and Mn2+ bound to the high-affinity site, Appl. Magn. Reson., 2003, 23, 571. 

D.I. Sanchez Machado, B. Chavira Willys, J. Lopez Cervantes, High Performance Liquid Chromatography with Fluorescence Detection for Quantitation of Tryptophan and Tyrosine in a Shrimp Waste Protein Concentrate, Journal of Chromatography, B, 863(1), 88 93 (2008). 

Sites at which nonsense mutations are detected are identified by their amber (A) or ochre (0) alleles (Coulondre and Miller, 1977). The 8 tyrosine codons in lacl each have two nonsense alleles, one amber and one ochre. The amber alleles at these sites are marked by the symbols and . 

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