Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:1.14.16.2 (tyrosine hydroxylase)
 14,760 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Neurocatin, a neuroregulatory factor isolated from mammalian brain, is a powerful affector of dopamine synthesis in striatal rat synaptosomes. Incubation of intact synaptosomes with neurocatin caused an increase in the rate of dopamine synthesis measured by accumulation of DOPA. The increase is rapid (within two minutes) and dependent on the concentration of added neurocatin. The stimulatory effect of neurocatin on dopamine synthesis occurred only in intact synaptosomes and was almost completely abolished by lysis of the synaptosomes with Triton X-100 or sonification prior to neurocatin addition. The kinetic parameters of tyrosine hydroxylase were measured in lysates prepared from synaptosomes preincubated with neurocatin. These showed that with increasing neurocatin concentration there was an increase in Vmax with no significant change in KM for the pteridine cofactor, compared to control. Activation of tyrosine hydroxylase by neurocatin is at least partially caused by a receptor mediated increase in phosphorylation of the enzyme. Protein kinase C and protein kinase II may be involved in this process.
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PMID:Activation of striatal tyrosine hydroxylase by neurocatin, a neuroregulator from mammalian brain. 135 63

Intact secretory granules isolated from bovine adrenal medulla express tyrosine hydroxylase (TH) activity. Granule-associated TH sediments on continuous sucrose gradients with dopamine beta-hydroxylase, a marker for granule membranes, indicating that TH is associated with chromaffin granules. Membranes prepared from lysed granules retain TH, whereas granule contents are free of the enzyme. TH immunoreactivity was detected in granule membranes by immunoblot analysis using a polyclonal antiserum against TH. TH immunoreactivity cannot be removed from membranes by washes in high ionic strength buffers and is only partially removed from membranes by treatment with either urea or Na2CO3. TH can be removed from granule membranes by the detergents Nonidet P-40, Triton X-100, and 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate. Treatment of membranes with a phosphatidylinositol-specific phospholipase C did not remove TH, ruling out the possibility of a glycosyl phosphatidyl anchor. Fractionation of granule membranes by temperature-induced phase separation in Triton X-114 revealed that TH is recovered in phases in which integral (detergent phase) and hydrophobic (phospholipid phase) membrane proteins are typically found. By contrast, TH from adrenal cytosol fractionated exclusively into the aqueous phase along with other soluble proteins. Digestion of granules with various protease enzymes revealed that TH is resistant to degradation, suggesting that the enzyme is embedded within membranes. TH becomes phosphorylated when intact granules are exposed to the catalytic subunit of the cAMP-dependent protein kinase, indicating that at least the N-terminal region of TH is exposed on the cytoplasmic surface of granules. These results establish that a fraction of TH is an integral component of bovine granule membranes. The association of TH with granule membranes may play a role in coordinating TH activity and catecholamine release.
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PMID:Tyrosine hydroxylase in secretory granules from bovine adrenal medulla. Evidence for an integral membrane form. 196 7

Peripheral afferent denervation (deafferentation) of the rodent main olfactory bulb produces a marked decrease in tyrosine hydroxylase (TH) activity and immunoreactivity in a population of juxtaglomerular dopaminergic neurons. Preservation of activity and immunostaining for aromatic L-amino acid decarboxylase implies that these cells do not die, but change phenotype. We now report that the steady-state level of TH mRNA markedly decreases in the adult mouse olfactory bulb in response to deafferentation. This reduction is permanent following intranasal irrigation with 0.17 M zinc sulphate (ZnSO4) but reversible following deafferentation produced by intranasal irrigation with 0.7% Triton X-100. The initial declines in TH activity, protein and mRNA of dopaminergic juxtaglomerular neurons observed after Triton X-100 treatment are all reversible as the steady-state level of TH mRNA gradually returns to control levels. Steady-state levels of mRNA for olfactory marker protein (OMP), a protein found in high concentrations in olfactory receptor neurons and their processes which innervate the olfactory bulb, were also monitored following deafferentation. Following treatment with either ZnSO4 or Triton X-100, the pattern of changes in steady-state levels of OMP mRNA was similar to that observed for TH. The steady-state level of PEP19 mRNA, a peptide previously localized to granule cells in the olfactory bulb, was not altered by deafferentation. These data indicate selective and parallel regulation of TH and OMP message and protein levels following deafferentation.
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PMID:Transneuronal regulation of neuronal specific gene expression in the mouse olfactory bulb. 197 Oct 84

The cellular localization of carbonic anhydrase (CAH) in the carotid body of the rat was investigated by means of Hansson's cobalt-precipitation technique in cultures of dissociated cells. In both young (2-day-old) and old (77-day-old) cultures, the parenchymal glomus (type-I) cells were selectively stained by this technique, and in addition expressed tyrosine hydroxylase and neuron-specific enolase as revealed by immunofluorescence. Enzymic reaction product of CAH appeared to be predominantly intracellular since staining was more intense and occurred more rapidly following permeabilization of the cell membranes with Triton X-100; its formation was inhibited by the CAH-inhibitor acetazolamide (1-10 microM) or by increasing the pH from 5.8 to 7.5. Cryostat sections of the carotid bifurcation revealed intense CAH-reaction product in cell clusters of the carotid body, in a few cells of the nodose ganglion, and in red blood cells. Neuronal cell bodies of the petrosal ganglion and superior cervical ganglion (SCG) were largely non-reactive. The SCG is known to contain clusters of small intensely fluorescent (SIF) cells, which were also non-reactive when grown in dissociated cell culture. Thus, although glomus and SIF cells are often considered to be similar cell types, functional CAH-activity appears unique to glomus cells, and this may be important for the physiological response of the carotid body to certain chemosensory stimuli.
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PMID:Carbonic anhydrase and neuronal enzymes in cultured glomus cells of the carotid body of the rat. 197 81

Subjecting either P2 fraction or purified synaptosomes isolated from rat brain to periods of anoxic incubation at 30 degrees resulted in activation of dopamine synthesis from tyrosine. This activation was approximately 2.5-fold when the anoxic incubation was carried out at pH 6.2 but was not significant when the pH was 7.4. Measurements of the tyrosine hydroxylase activity at pH 6.2 in Triton X-100-treated preparations of the P2 fraction showed that, after 20 min of anaerobiosis, the Km for pterine cofactor decreased by 39% and the Ki for dopamine increased by 44%; there was no change in the Km for tyrosine. Half-maximal activation of dopamine synthesis occurred in 10 min of anaerobic incubation, and the reversal upon addition of oxygen had a half-time of 15 min. Addition of forskolin or dibutyryl cyclic AMP to anaerobic incubations of P2 fraction did not result in significant activation of dopamine synthesis. Either the removal of calcium or the addition of calmodulin inhibitor, trifluoperazine, substantially decreased the activation of dopamine synthesis induced by periods of anaerobiosis. It appears that during anoxic incubation tyrosine hydroxylase underwent an activation which occurred over a period of minutes, was stable to detergent treatment, and was fully reversed over a period of minutes following reoxygenation. This activation was, at least in part, dependent on the presence of calcium and was sensitive to the calmodulin antagonist trifluoperazine.
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PMID:Activation of tyrosine hydroxylase in the central nervous system by anaerobiosis. 286 73

Tyrosinase activity increased in Cloudman S-91 mouse melanoma cell homogenates incubated at 37 degrees C for a minimum of 8 h. Enzyme activity continued to increase for 48 h at which time the maximal level of activation was observed. Activation did not occur at 4 degrees C and did not occur in the cytosol fraction of the cell, suggesting that the response was localized to melanosomes. The activated enzyme was resistant to solubilization with the nonionic detergent, Triton X-100, and preparation of homogenates in this detergent did not inhibit the temperature-dependent activation of the melanosomal fraction of the cell. The activation process increased the Vmax of tyrosinase 10-fold and lowered the Km by a factor of 2 as determined by the tyrosine hydroxylase assay. The increase in tyrosinase activity was detectable by three assay methods: tyrosine hydroxylation, melanin synthesis, and by tyrosine decarboxylation. The formation of melanin, however, was found to be 1/20 that of either tyrosine hydroxylation or decarboxylation, a finding which suggests that the melanin pathway may be blocked at 5,6-dihydroxyindole. The "self-activation" response could not be mimicked by incubating cell homogenates with cyclic AMP-dependent protein kinase. Activated tyrosinase could be inhibited by the addition of fresh cell extracts, a finding which suggests that tyrosinase inhibitors may be present in these cells.
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PMID:Activation of tyrosinase in mouse melanoma cell cultures. 288 48

A previous published assay method for tyrosine hydroxylase by the evolution of 14CO2 was modified to a two-step procedure to allow reliable measurement of large numbers of samples containing low tyrosine hydroxylase activity. The reliability of the method was examined in detail. Properties of rat brain and pineal tyrosine hydroxylase solubilized with 0.2% Triton X-100 were as follows. The apparent Km values of the brain enzyme for L-tyrosine with 1 mM-(6-DL)-5,6,7,8-tetrahydro-L-erythro-biopterin (BPH4) as cofactor and for BPH4 with 62 microM-L-tyrosine as substrate were approximately 25 microM and 85 microM, respectively. The Km's for L-tyrosine with 1 mM-(6-DL)-5,6,7,8-tetrahydro-6-methylpterin (6MPH4) as cofactor and for 6MPH4 with 210 microM-L-tyrosine as substrate were 68 microM and 270 microM, respectively. The marked substrate inhibition by high concentrations of L-tyrosine was observed only when BPH4 was used as cofactor. High concentrations of BPH4 inhibited the reaction slightly. The kinetic properties of tyrosine hydroxylase in the pineal extract were similar to those of the brain enzyme, except that a Lineweaver-Burk plot of reciprocal velocity versus the reciprocal concentration of BPH4 with 62 microM-L-tyrosine as substrate deviated downward at a BPH4 concentration of about 100 microM. Analyses of the plot indicated that the peculiar kinetic property may represent either the reaction occurring at two independent sites or with two forms (6L- and 6D-isomers) of the tetrahydrobiopterin cofactor, with apparent Km for BPH4 of 23 microM and 1025 microM, respectively, or the negatively cooperative ligand binding with a Hill coefficient of 0.72. Based on the results obtained as reported above the standard assay conditions of tyrosine hydroxylase in tissue extracts were established. Using the assay method and conditions, the absence of the daily rhythmicity of tyrosine hydroxylase in rat pineal glands and three discrete brain areas was demonstrated. The findings, especially on pineal tyrosine hydroxylase, are discussed in relation to the daily change of noradrenaline turnover.
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PMID:The activity of rat pineal and brain tyrosine hydroxylase during the daily cycle of light and darkness as determined by the modified 14CO2 assay method. 610 56

Peripheral afferent input regulates the expression of dopaminergic properties in a population of local circuit intrinsic neurons of the rodent olfactory bulb. Lesions of the olfactory receptor neurons produced in the mouse by intranasal irrigation with either ZnSO4 or Triton X-100 and in the rat by surgical deafferentation or axotomy are associated with a decrease in the levels of dopamine (DA), the DA metabolite 3,4-dihydroxyphenylacetic acid (DOPAC), the activity of the enzyme tyrosine hydroxylase (TH), bulb weight and an increase in norepinephrine (NE) levels in the olfactory bulb. The anatomical correlates of the biochemical sequelae of deafferentation of olfactory bulb DA neurons were studied using immunohistochemical techniques to localize TH. Within 3 to 4 weeks all lesions produced a dramatic and uniform reduction in TH staining of the juxtaglomerular DA neurons and their processes which was paralleled by a reduction in DA and DOPAC levels and bulb weight. Seven weeks following reversible chemical lesions produced by Triton X-100, DA and DOPAC levels and tissue weight as well as TH staining in the juxtaglomerular neurons returned to control levels. These observations suggested that DA neurons remained present even when not demonstrable with TH antibodies. Additional evidence for the continued presence of the DA neurons was the ability of the olfactory bulbs from both lesioned mouse and rat to synthesize DA from exogenously administered L-3,4-dihydroxyphenylalanine (l-DOPA). These data suggested that the decrease in DA levels and TH staining in the olfactory bulb following lesions of the olfactory receptor neurons were produced by transneuronal mechanisms since there was no direct injury of the bulb. Furthermore, the demonstration that following reinnervation, catecholamine synthetic capacity is restored suggests that the juxtaglomerular dopamine neurons remain in the bulb and that afferent receptor input is required for expression of TH enzyme.
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PMID:Transneuronal regulation of tyrosine hydroxylase expression in olfactory bulb of mouse and rat. 613 Jan 33

Lumbosacral (L6-S1) spinal cord neurons in the cat were retrogradely labelled after uptake of horseradish peroxidase by their severed axons in the upper cervical (C3-C4) dorsolateral funiculus. Sections of L6-S1 containing labelled neurons were then processed immunocytochemically using antibodies against dopamine-beta-hydroxylase or tyrosine hydroxylase, two enzymes responsible for the synthesis of catecholamines. Two hundred and ninety eight retrogradely-labelled cells within laminae III-V of the dorsal horn were examined under high power (x 1000) with the light microscope. In Triton X-100-treated material, only 13% of these cells had catecholamine-containing varicosities closely apposed to their somata and proximal dendrites, which suggests that in comparison with the postsynaptic dorsal column pathway, spinocervical tract neurons are only sparsely innervated by descending catecholaminergic axons.
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PMID:Relationships between spinocervical tract neurons and descending catecholamine-containing axons in the cat. 791 39

Tyrosinase, tyrosinase-related protein-1 (TRP-1), and tyrosinase-related protein-2, (TRP-2, dopachrome tautomerase) were shown by immunoblotting and enzyme assays to copurify from extracts of Cloudman S91 melanoma cells. Antibodies to TRP-1 and TRP-2 immunoprecipitated tyrosinase activity, suggesting a stable interaction (complex) among these proteins. The tyrosine hydroxylase activity of tyrosinase was reduced in the complexed form; treatment with Triton X-100 dissociated the complex and activated the tyrosinase present within it. To further study this complex, we employed sucrose gradient density centrifugation of extracts from cultured murine melanocytes. Tyrosinase, TRP-1, and TRP-2 all existed in high molecular weight "multimers" of approximately 200 to > 700 kilodaltons. Extraction of cells with buffers containing the detergent CHAPS preserved the high molecular weight multimers; Triton X-100 caused their dissociation into monomers. Low pH, low ionic strength, and millimolar concentrations of calcium ions favored the maintenance of multimers. The results of this study demonstrate that the participation of tyrosinase, TRP-1, and TRP-2 in a multimeric complex could have important physiologic consequences, and raise the possibility that some of the well-known interactions between coat color genes may be explained by intermolecular interactions between the gene products.
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PMID:High-molecular-weight forms of tyrosinase and the tyrosinase-related proteins: evidence for a melanogenic complex. 804 Jun 9


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