Expression of four types of human tyrosine hydroxylase in COS cells

Kobayashi, Kazuto; Kiuchi, Kazutoshi; Ishii, Akira; Kaneda, Norio; Kurosawa, Yoshikazu; Fujita, Kazuo; Nagatsu, Toshiharu

doi:10.1016/0014-5793(88)80526-x

Cited by 39 publications

(12 citation statements)

References 28 publications

(15 reference statements)

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“…Only one of the two forms was stimulated by Fe(I1) [half-maximal activation at about 20 pM Fe(II)]. Moreover, the same authors previously reported that of the four isozymes, transiently expressed in COS cells, only TH2 was activated by Fe(I1) [16]. Our results on the purified recombinant human isozymes are clearly different.…”

Section: Discussioncontrasting

confidence: 47%

Recombinant human tyrosine hydroxylase isozymes

Haavik

Bourdellès

Martı́nez

et al. 1991

European Journal of Biochemistry

109

131

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Human tyrosine 3-monooxygenase (tyrosine hydroxylase) exists as four different isozymes (TH1-TH4), generated by alternative splicing of pre-mRNA. Recombinant TH1, TH2 and TH4 were expressed in high yield in Escherichiu coli. The purified isozymes revealed high catalytic activity [when reconstituted with Fe(II)] and stability at neutral pH. The isozymes as isolated contained 0.04-0.1 atom iron and 0.02-0.06 atom zinc/enzyme subunit. All three isozymes were rapidly activated (1 3 -40-fold) by incubation with Fe(I1) salts (concentration of iron at half-maximal activation = 6-14 pM), and were inhibited by other divalent metal ions, e.g. Zn(II), Co(I1) and Ni(I1). They all bind stoichiometric amounts of Fe(I1) and Zn(I1) with high affinity (Kd = 0.2-3 pM at pH 5.4-6.5). Similar time courses were observed for binding of Fe(I1) and enzyme activation. In the absence of any free Fe(I1) or Zn(II), the metal ions were released from the reconstituted isozymes. The dissociation was favoured by acidic pH, as well as by the presence of metal chelators and dithiothreitol. The potency of metal chelators to remove iron from the hydroxylase correlated with their ability to inhibit the enzyme activity. These studies show that tyrosine hydroxylase binds iron reversibly and that its catalytic activity is strictly dependent on the presence of this metal.Tyrosine 3-monooxygenase (tyrosine hydroxylase) is a tetrahydropteridine-dependent enzyme which catalyses the rate-limiting step in the biosynthesis of catecholamines [ 11. Early studies showed that the enzyme activity is stimulated by ferrous iron and inhibited by certain iron chelators [2 -51.Subsequently, it was shown that the enzyme isolated from various tissues contained significant amounts of iron [6 -81, which seems to be involved in oxygen activation, an essential step in the catalytic reaction [6-81. However, partly due to the difficulties involved in obtaining pure tyrosine hydroxylase, no systematic study has been performed in order to determine the metal-binding properties of this enzyme.The recent expression of cloned tyrosine hydroxylase in high yield in eukaryotic cells using the baculovirus system [9, 101 and in prokaryotic cells [ 10 a] has, however, changed this situation. Human tyrosine hydroxylase exists as four different isozymes (TH1-TH4), generated through alternative splicing of pre-mRNA [ l l -131. Since these isozymes have different amino acid sequences in their regulatory N-terminal region and different distribution in the nervous system [14], it has been postulated that the occurrence of multiple molecular forms of the enzyme has important functional consequences. Previous studies, in which the isozymes have been expressed in low quantities in eukaryotic systems, have also demonstrated different kinetic properties of the various isozymes [15, 161. Correspondence to J. Haavik, Department of Biochemistry, UniAbbreviations. TH 1 -TH4, tyrosine hydroxylase isozymes 1 -4. Enzyme. Tyrosine 3-monooxygenase or tyrosine hydroxylase versity of Bergen, N-5009 Berge...

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Section: Discussioncontrasting

confidence: 47%

Recombinant human tyrosine hydroxylase isozymes

Haavik

Bourdellès

Martı́nez

et al. 1991

European Journal of Biochemistry

109

131

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“…Human TH yields immunoreactive bands of 62 to 68 kD [40, 41], whereas that from the rat is estimated at 60 kD [42]. Our Western blot results showed that TH protein was not detected in HUMSCs treated with NCM only.…”

Section: Resultsmentioning

confidence: 83%

Conversion of Human Umbilical Cord Mesenchymal Stem Cells in Wharton's Jelly to Dopaminergic Neurons In Vitro: Potential Therapeutic Application for Parkinsonism

Fu¹,

Cheng²,

et al. 2005

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Human mesenchymal stem cells isolated from Wharton's jelly of the umbilical cord were induced to transform into dopaminergic neurons in vitro through stepwise culturing in neuron-conditioned medium, sonic hedgehog, and FGF8. The success rate was 12.7%, as characterized by positive staining for tyrosine hydroxylase (TH), the rate-limiting catecholaminergic synthesizing enzyme, and dopamine being released into the culture medium. Transplantation of such cells into the striatum of rats previously made Parkinsonian by unilateral striatal lesioning with the dopaminergic neurotoxin 6-hydroxydopamine partially corrected the lesion-induced amphetamine-evoked rotation. Viability of the transplanted cells at least 4 months after transplantation was identified by positive TH staining and migration of 1.4 mm both rostrally and caudally. These results suggest that human umbilical mesenchymal stem cells have the potential for treatment of Parkinson's disease. STEM CELLS 2006; 24:115-124

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“…TH, DBH, and ChAT genes could be either primary target genes of RA receptors or indirectly modulated by RA. The characterization of TH, DBH, and ChAT promoter elements (Lewis et al, 1987;Kobayashi et al, 1989;Ibanez and Persson, 1991;Bejanin et al, 1992;Shaskus et al, 1992) will now allow these possibilities to be tested by searching for RA receptor binding sites in the transcriptional control regions of the three genes.…”

Section: Discussionmentioning

confidence: 97%

Retinoic acid induces cholinergic differentiation of cultured newborn rat sympathetic neurons

Berrard

Biguet

Houhou

et al. 1993

J of Neuroscience Research

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Many studies provide evidence that retinoic acid (RA), an endogenous derivative of vitamin A, plays a role in the development of the nervous system. We now report that RA controls the neurotransmitter phenotype of post-mitotic rat sympathetic neurons in cell culture. RA added to the culture medium increased the specific activity of choline acetyltransferase (ChAT) and the level of acetylcholine (ACh). Concomitantly, RA reduced the specific activities of two catecholamine synthetic enzymes, tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH) and the level of norepinephrine (NE). After a 2 week treatment with 5 microM RA, ChAT was increased by 5-10 fold, whereas TH and DBH were decreased by 10-15 fold and 2-3 fold, respectively, as compared to sympathetic neurons grown in the absence of RA. The modulation of the activity of the three enzymes was dose-dependent and followed a similar time course. The decrease of TH expression was demonstrated to be due to a decreased number of TH molecules.

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Expression of four types of human tyrosine hydroxylase in COS cells

Cited by 39 publications

References 28 publications

Recombinant human tyrosine hydroxylase isozymes

Recombinant human tyrosine hydroxylase isozymes

Conversion of Human Umbilical Cord Mesenchymal Stem Cells in Wharton's Jelly to Dopaminergic Neurons In Vitro: Potential Therapeutic Application for Parkinsonism

Retinoic acid induces cholinergic differentiation of cultured newborn rat sympathetic neurons

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