A structural approach into human tryptophan hydroxylase and its implications for the regulation of serotonin biosynthesis

Tryptophan hydroxylase (TPH) catalyzes the 5-hydroxylation of tryptophan, which is the first step in the biosynthesis of indoleamines (serotonin and melatonin). Serotonin functions mainly as a neurotransmitter, whereas melatonin is the principal hormone secreted by the pineal gland. TPH belongs to the family of the aromatic amino acid hydroxylases, including phenylalanine hydroxylase (PAH) and tyrosine hydroxylase (TH), which all have a strict requirement for dioxygen, non-heme iron (II) and tetrahydrobiopterin (BH4). During the last three years there has been a formidable increase in the amount of structural information about PAH and TH, which has provided new insights into the active site structure, the binding of substrates, inhibitors and pterins, as well as on the effect of disease-causing mutations in these hydroxylases. Although structural information about TPH is not yet available, the high sequence homology between the three mammalian hydroxylases, notably at the catalytic domains, and the similarity of the reactions that they catalyze, indicate that they share a similar 3D-structure and a common catalytic mechanism. Thus, we have prepared a model of the structure of TPH based on the crystal structures of TH and PAH. This structural model provides a frame for understanding the specific interactions of TPH with L-tryptophan and substrate analogues, BH4 and cofactor analogues, L-DOPA and catecholamines. The interactions of these ligands with the enzyme are discussed focusing on the physiological and pharmacological regulation of serotonin biosynthesis, notably by tryptophan supplementation therapy and substitution therapy with tetrahydrobiopterin analogues (positive effects), as well as the effect of catecholamines on TPH activity in L-DOPA treated Parkinson's disease patients (enzyme inhibition).     

Tetrahydrobiopterin binding to aromatic amino acid hydroxylases. Ligand recognition and specificity

The three aromatic amino acid hydroxylases (phenylalanine, tyrosine, and tryptophan hydroxylase) and nitric oxide synthase (NOS) all utilize (6R)-l-erythro-5,6,7,8-tetrahydrobiopterin (BH(4)) as cofactor. The pterin binding site in the three hydroxylases is well conserved and different from the binding site in NOS. The structures of phenylalanine hydroxylase (PAH) and of NOS in complex with BH(4) are still the only crystal structures available for the reduced cofactor-enzyme complexes. We have studied the enzyme-bound and free conformations of BH(4) by NMR spectroscopy and molecular docking into the active site of the three hydroxylases, using endothelial NOS as a comparative probe. We have found that the dihydroxypropyl side chain of BH(4) adopts different conformations depending on which hydroxylase it interacts with. All the bound conformations are different from that of BH(4) free in solution at neutral pH. The different bound conformations appear to result from specific interactions with nonconserved amino acids at the BH(4) binding sites of the hydroxylases, notably the stretch 248-251 (numeration in PAH) and the residue corresponding to Ala322 in PAH, i.e., Ser in TH and Ala in TPH. On the basis of analysis of molecular interaction fields, we discuss the selectivity determinants for each hydroxylase and explain the high-affinity inhibitory effect of 7-tetrahydrobiopterin specifically for PAH.     

芳香族氨基酸对沼泽红假单胞菌合成CoQ10的影响

CoQ10具有呼吸链电子传递者、抗氧化性、调控基因表达等多种生理生化功能.目前不仅用作药物也用作食品添加剂.微生物发酵法是当前生产CoQ10的主要方法.在细菌中,芳香族氨基酸和CoQ10的苯核环都是通过莽草酸途径合成;它们在生物体中各自的合成存在着相互调控作用.本试验通过在培养过程中添加芳香族氨基酸来考察了其对沼泽红假单胞菌J001合成CoQ10的影响.结果表明:当色氨酸添加量≥15 mg/L时对CoQ10的合成具有强列的抑制作用;苯丙氨酸添加量≥50mg/L时对CoQ10的合成具有一定的抑制作用但当添加量≥75mg/L后对CoQ10的合成变为一定的促进作用;酪氨酸添加量为75～175 mg/L时对CoQ10的合成具有一定的促进作用,但当添加量≥200 mg/L后对CoQ10的合成变为一定的抑制作用.当添加苯丙氨酸100 mg/L+酪氨酸150mg/L时,CoQ10含量达最高(25.6±1.3 mg CoQ10/g干细胞),比不添加对照提高52.2%.以上结果说明:该菌株CoQ10的合成受到3种芳香族氨基酸的调控,3-脱氧-D-阿拉伯己酮糖-7-磷酸合成酶是同工酶系统并主要受色氨酸调控,无色氨酸培养基及在培养期间添加适量的苯丙氨酸与酪氨酸对该菌株合成CoQl.有利.     

Synthetic analogues of tetrahydrobiopterin with cofactor activity for aromatic amino acid hydroxylases

Tetrahydrobiopterin (THB) analogues with 6-alkoxymethyl substituents, 3a-j, where the substituents were straight- and branched-chain alkyl ranging from methyl to octyl, have been synthesized by the Taylor method from pyrazine ortho amino nitriles by guanidine cyclization, hydrolysis in aqueous NaOH, and catalytic hydrogenation over Pt in trifluoroacetic acid (TFA). The best of these compounds, 3b, is an excellent cofactor for phenylalanine hydroxylase, tyrosine hydroxylase (V = 154% of THB), and tryptophan hydroxylase, does not destablize the binding of substrate (Kmtyr = 23 microM), and is recycled by dihydropteridine reductase (V = 419% of THB). The compounds are being evaluated as cofactor replacements in biopterin-deficiency diseases.     

Effects of chronic monoamine oxidase inhibitor treatment on biogenic amine metabolism in rat brain.

The effects of acute and chronic administration of phenelzine and tranylcypromine on rat brain monoamine metabolism have been examined. Peak increases in norepinephrine, dopamine and 5-hydroxytryptamine occurred between 1 and 7 days with monoamine oxidase inhibitor treatments followed by a gradual decline in brain monoamines towards control levels with continued chronic drug administration. There was an associated adaptive increase in tryptophan hydroxylase but no change in tyrosine hydroxylase activity with chronic phenelzine treatment. Tranylcypromine did not affect tryptophan hydroxylase or tyrosine hydroxylase activities but was associated with a significant increase in aromatic amino acid decarboxylase activity after 14 and 21 days of treatment.     

Studies on α-glucosidase inhibitors development: magic molecules for the treatment of carbohydrate mediated diseases

α-Glucosidase (EC 3.2.1.20) enzyme belongs to the glycosidase family enzymes, cleave the glycosidic bond of the oligosaccharides that liberate glucose and its inhibition retards the carbohydrate digestion. In the present review, we have discussed the structural features of different α-glucosidase inhibitors (small molecules) responsible for the inhibitory activities. The reported computational studies including, QSAR, pharmacophore modelling, homology models, docking (with analogs enzymes), etc revealed that the topological, electronic and hydrophobicity properties determine the interactions of those molecules. The aromatic substituents connected with flexible bonds in the molecules have significant effect on the interactions, which may due to the presence of aromatic amino acid residues in the active site. The reported homology modelled and other analogs enzymes (enzymes of other species) also confirmed the existence of aromatic residue (amino acids) especially, histidine, phenylalanine and tyrosine in their active site along with the polar (glutamic and aspartic acids) residues. Multiple sequence alignments of the α-glucosidase enzymes (from different species) described that the abovementioned amino acid residues are present in the active site of all the studied enzymes. Recently, Celgosivir (MIGENIX Inc) is an oral prodrug of the natural product castanospermine used for the treatment of HCV infection by inhibiting α-glucosidase I. BMN-701 is an α-glucosidase inhibitors in the phase I pipeline (BioMarine) for the treatment of Pompe diseases. CKD-711 and CKD-711a are aminooligosaccharide α-glucosidase inhibitors and the in vitro study of CKD-711 showed similar effects to acarbose on porcine intestinal maltase and sucrase (IC50s of 2.5 and 0.5 μg/ml). This review also concluded that many α-glucosidases inhibitors obtained from natural products are used for the treatment of various carbohydrate mediated diseases. The structural analysis of these synthetic and natural derivatives guide for the development of novel semisynthetic/synthetic α-glucosidase inhibitors with free of toxicities.     

急性色氨酸、苯丙氨酸和酪氨酸剥夺对酒依赖者线索暴露诱导的饮酒迫促性的作用

目的:评价急性色氨酸、苯丙氨酸和酪氨酸剥夺(CMD)对酒依赖者线索暴露诱导的饮酒迫促性的作用。方法:采用随机双盲交叉对照的方法,选择符合ICD-10和DSM-Ⅳ诊断标准的男性酒依赖患者12名,并排除任一符合DSM-Ⅳ轴Ⅰ诊断的其他精神障碍。在签署知情同意书的情况下,每位受试者间隔至少7d分别在实验当天禁食的情况下给予含人体必需的16种氨基酸的氨基酸饮料(BLANCE)或缺失色氨酸、苯丙氨酸和酪氨酸的氨基酸饮料(CMD),实验的前一天低蛋白饮食,实验当天结束后给予高蛋白饮食。实验当天,在给予氨基酸饮料前后进行Tiffany饮酒迫促性量表、焦虑可视模拟标尺、抑郁可视模拟标尺、副反应量表测查以及血压、心率检测。给予氨基酸饮料6h后进行非酒精(纯净水)相关线索和酒精(白酒)相关线索暴露,并评价暴露前后饮酒迫促性、焦虑可视模拟标尺分、抑郁可视模拟标尺分和血压、心率的变化。结果:CMD组在非酒精相关线索和酒精相关线索暴露后饮酒意愿(Intention/desire to drinking)和舒张压均增加,差异具有显著性(P=0.0217,0.038),而BLANCE组在非酒精相关线索和酒精相关线索暴露后饮酒意愿、舒张压差异均无显著性。焦虑可视模拟标尺分、抑郁可视模拟标尺分、收缩压、心率在两组中的非酒精相关线索和酒精相关线索暴露前后的差异无显著性。在安全性方面,所有受试者中有1名出现腹泻,但是病人尚能耐受。其余少见的副反应主要有恶心、头晕、困倦等,皆为轻度或中度。结论:急性色氨酸、苯丙氨酸和酪氨酸剥夺(CMD)后酒依赖患者在非酒精和酒精相关线索暴露后的饮酒意愿增加,舒张压增高,但是对焦虑和抑郁情绪没有影响。口服氨基酸饮料的副反应少且轻微。     

Sapropterin dihydrochloride, 6-R-L-erythro-5,6,7,8-tetrahydrobiopterin, in the treatment of phenylketonuria

Sapropterin dihydrochloride, 6-R-L-erythro-5,6,7,8-tetrahydrobiopterin (BH4) is being introduced in the US for treatment of phenylketonuria (PKU). This compound has been in use in Europe to treat mild forms of PKU. Tetrahydrobiopterin is the cofactor in the hydroxylation reaction of the three aromatic amino acids phenylalanine, tyrosine and tryptophan. It is also involved in other reactions, which are not the focus of this review. The cofactor BH4 is synthesized in many tissues in the body. The pathway of BH4 biosynthesis is complex, and begins with guanosine triphosphate (GTP). The first reaction that commits GTP to form pterins is GTP cyclohydrolase. Several reactions follow resulting in the active cofactor BH4. During the hydroxylation reaction BH4 is oxidized to quinonoid-BH2, which is recycled by dihydropteridine reductase, resulting in the active cofactor. It was discovered that some patients with PKU had a decline in blood phenylalanine after oral intake of BH4. This response to BH4 is not the result of change in the synthesis or regeneration of the cofactor, but rather an effect on the mutant enzyme phenylalanine hydroxylase either by accommodating the higher K(m) of the mutant enzyme or by acting as a chaperone for the mutant enzyme. This response has become of intense interest in the treatment of PKU.     

偏振荧光光度法同时测定氨基酸注射液及动、植物浸出液中酪氨酸、色氨酸的研究

酪氨酸、色氨酸和苯丙氨酸具有荧光,本文使用互相垂直的偏振片有效地消除了苯丙氨酸的干扰和溶剂散射光的影响;附加滤光片,进一步提高了酪氨酸与色氨酸荧光测定的选择性。线性范围分别为0.02～12.0mg·L^-1(酪氨酸)与0.01～2.5mg·L^-1(色氨酸)。此法可直接用于氨基酸注射液和动、植物浸出液中游离酪氨酸、色氨酸的分析,结果满意。     

Effect of antidepressant drugs on monoamine synthesis in brain in vivo.

The activity of tryptophan and tyrosine hydroxylase were estimated in vivo by measuring the accumulation during 30 min of 5-hydroxytryptophan (5-HTP) and 3,4-dihydroxyphenylalanine (DOPA), respectively, after inhibition of aromatic amino acid decarboxylase by administration of m-hydroxybenzylhydrazine (NSD 1015) (100 mg/kg, i.p.). Whereas the activity of tyrosine hydroxylase in the dopamine-rich striatum was sensitive to haloperidol, which caused a significant increase in accumulation of DOPA, there was no effect of haloperidol in the predominantly noradrenergic frontoparietal cortex, confirming that the activity of tyrosine hydroxylase, measured in the frontoparietal cortex, is essentially localized in noradrenergic neurones. In the frontoparietal cortex of the rat the in vivo activity of tryptophan and tyrosine hydroxylase were equipotently attenuated by imipramine, while the selective blocker of the uptake of noradrenaline, desipramine and the selective blocker of the uptake of serotonin, citalopram, reduced only tyrosine or tyrosine hydroxylase respectively. Milnacipran, an antidepressant which inhibits the uptake of both monoamines to a similar extent, decreased the synthesis of both monoamines equipotently. The monoamine oxidase inhibitor, clorgyline, also reduced the synthesis of both monoamines. Thus, the in vivo inhibition of the synthesis of monoamines would appear to be mediated by an increase in synaptic concentration of monoamines, resulting from the inhibition of the uptake or catabolism of monoamines. Chronic administration of citalopram led to a significant increase of the basal synthesis of 5-hydroxytryptamine (5-HT). Milnacipran, given chronically, significantly enhanced the basal synthesis of both 5-HT and noradrenaline (NA).(ABSTRACT TRUNCATED AT 250 WORDS)     

Determination of the ratios of the aromatic amino acid residues by first- or second-derivative UV spectrometry for a simple characterization of peptides

A method for the simultaneous determination of the ratios of the three aromatic amino-acid residues in peptides was set up in acidic conditions. Binary and ternary mixtures of these amino acids were prepared, and first- and second-derivative spectra then calculated from their 0.1 nm resolution spectra between 240 and 320 nm. Certain spectral bands were chosen to differentiate tyrosine from tryptophan on the first-derivative spectra, and phenylalanine from tyrosine and tryptophan on the second-derivative spectra. Variation of the amplitude of the chosen bands was shown to be a linear function of the ratio of the aromatic amino acids in the mixture. This technique was validated with peptides whose sequence was known. The difference between theoretical and experimentally determined ratios was lower than 10%. Since the results are obtained as ratios, neither the concentration nor the nature of the peptide has to be known. The feasibility of application using a photodiode array detector with high resolution in reversed-phase high-performance liquid chromatography is discussed. © Munksgaard 1995.     

Synthesis and biological properties of alpha-mono- and alpha-difluoromethyl derivatives of tryptophan and 5-hydroxytryptophan

The syntheses of alpha-mono- and alpha-difluoromethyl derivatives of tryptophan and 5-hydroxytryptophan are described. In an attempt to selectively regulate serotonin synthesis, alpha-(mono- and difluoromethyl)tryptophan were tested in vivo as precursors (or prodrugs) of their 5-hydroxy analogues. Although alpha-(mono- and difluoromethyl)-5-hydroxytryptophans are potent irreversible inhibitors of aromatic amino acid decarboxylase (equipotent to alpha-difluoromethyl-Dopa), only alpha-(monofluoromethyl)tryptophan affects the level of serotonin in vivo (small decrease), alpha-(difluoromethyl)tryptophan being a very poor substrate of the activating (or helper) enzyme, tryptophan hydroxylase.     

Structural and mechanistic implications of incorporating naturally occurring aberrant mutations of human dihydropteridine reductase into a rat model

Phenylketonuria (PKU) is a debilitating hereditary disorder related to an individual's inability to convert phenylalanine to its usual tyrosine product. The genetic errors occur in three regions: in the cooperative enzymes phenylalanine hydroxylase (PAH) and dihydropteridine reductase (DHPR), and in the biosynthetic pathway from GTP to the hydroxylation cofactor, tetrahydrobiopterin (BH₄). Many instances of naturally occurring defects in DHPR metabolism have been identified, and in most cases the error has been equated with an altered enzyme gene sequence. Using computer graphics, this report analyses the altered structural characteristics of eight of the enzymes encoded by mutant gene sequence and provides logical explanations for their diminished enzyme activities. In one instance, that of a threonine insertion, a mutant construct of the rat analog has been expressed in Escherichia coli and the DHPR isolated and characterised, confirming the marked changes this insert can create.     

CHAINS OF ALTERNATING SULFUR AND π-BONDED ATOMS IN EIGHT SMALL PROTEINS

This paper demonstrates the existence of regions in eight small globular proteins in which the side chains of sulfur-containing amino acids (cysteine and methionine) alternate in space with side chains of aromatic amino acids (histidine, phenylalanine, tryptophan and tyrosine). The proteins are: rubredoxin, high potential iron protein, cytochrome c, flavodoxin, deoxyhemoglobin, trypsin inhibitor, ribonuclease-S, and lysozyme. The sulfur-φ-bonded ‘chains’ involve a minimum of five and a maximum of 10 amino acids, and contain the most polarizable atoms within proteins. S-φ-chains give extra stability to the folding of proteins; they may also afford paths for the step-wise movement of electrons.     

Disorders of tetrahydrobiopterin metabolism and their treatment

Tetrahydrobiopterin (BH4) deficiencies are disorders affecting phenylalanine metabolism in liver and neurotransmitters biosynthesis in brain. BH4 is the essential cofactor in the enzymatic hydroxylation of 3 aromatic amino acids (phenylalanine, tyrosine, and tryptophan). BH4 is synthesized from guanosine triphosphate (GTP) catalyzed by GTP cyclohydrolase I (GTPCH), 6-pyruvoyl-tetrahydropterin synthase, and sepiapterin reductase (SPR), and in aromatic amino acids hydoxylating system is regenerated by pterin-4a-carbinolamine dehydratase (PCD) and dihydropteridine reductase (DHPR). To date, 4 enzyme deficiencies (GTPCH, PTPS, DHPR, PCD) have been reported and they all follow an autosomal recessive mode of inheritance. The incidence of BH4 deficiency is at 1 in 1,000,000, except that in Taiwanese (much higher than in Japanese and Caucasians). BH4 deficiency has been diagnosed in patients with hyperphenylalaninemia (HPA) by neonatal mass-screening based on BH4 oral loading tests, analysis of urinary or serum pteridines, and measurement of dihydropterindine reductase (DHPR) activity in blood from a Guthrie card. BH4 deficiency without treatment causes combined symptoms of HPA and neurotransmitter (dopamine, norepinephrine, epinephrine, and serotonin) deficiency, such as red hair, psychomotor retardation, and progressive neurological deterioration. Treatment of BH4 deficiencies consists of BH4 supplementation (2-20 mg/kg per day) or diet to control blood phenylalanine concentration and replacement therapy with neurotransmitters precausers (L-dopa/CarbiDOPA and 5-hydroxytryptophan), and supplements of folinic acid in DHPR deficiency.     

Brain biochemical and behavioral changes in 4 weeks old rats after neonatal oxygen deprivation.

The acquisition of a conditioned avoidance response (CAR) and the effects on monoamine neurotransmittor synthesis was investigated in 28 days old rats after neonatal oxygen deprivation (100% N₂ for 20 min or 6% O₂-94% N₂ for 4.5 hr). The rats subjected to 6% O₂-94% N₂ for 4.5 hr at 1 day of age were markedly inferior in the CAR acquisition than the control group. The animals subjected to 20 min neonatal anoxia did not differ in avoidance responding compared to controls. Tyrosine hydroxylase and tryptophan hydroxylase activity was studied in vivo in whole brain at 28 days of age by measuring the accumulation of dihydroxyphenylalanine (DOPA) and 5-hydroxytryptophan (5-HTP) respectively after inhibition of aromatic L-amino acid decarboxylase with NSD 1015. An inhibition of tyrosine hydroxylase and tryptophan hydroxylase activity was found after neonatal exposure to 6% O₂-94% N₂ for 4.5 hr while the rats exposed to 100% N₂ for 20 min did not differ from controls. The precursor amino acids tyrosine and tryptophan and the levels of the endogenous monoamines dopamine (DA), noradrenaline (NA) and 5-hydroxytryptamine (5-HT) did not differ from controls in any of the groups. It is suggested that the observed behavioral deficits after prolonged hypoxia may be due to an impaired development of central catecholamine mechanisms.     

An amino acid mixture deficient in phenylalanine and tyrosine reduces cerebrospinal fluid catecholamine metabolites and alcohol consumption in vervet monkeys

 An amino acid mixture devoid of tryptophan, given orally, was previously shown to reduce cerebrospinal fluid levels of tryptophan and 5-hydroxyindoleacetic acid in vervet monkeys, as compared to a control mixture containing all essential amino acids. In the present study, we tested the possibility that a similar amino acid mixture containing tryptophan, but devoid of phenylalanine and tyrosine (the amino acid precursors of catecholamine neurotransmitters), would influence dopamine and noradrenaline metabolism. Five hours after the administration of this mixture to vervet monkeys, cerebrospinal fluid levels of homovanillic acid and 3-methoxy-4-hydroxyphenylethylene glycol were reduced by 27.4% and 26.9%, respectively. Both effects were statistically significant. Plasma tyrosine (-30%) and the ratio of tyrosine to the sum of other large neutral amino acids (ΣLNAA) were also significantly reduced. The behavioral efficacy of phenylalanine/tyrosine depletion was compared with that of tryptophan depletion in a primate model of voluntary alcohol consumption. All three drinks lowered alcohol consumption, but the effects of the tryptophan-deficient amino acid mixture were not different from those of the balanced amino acid control. The phenylalanine/tyrosine-deficient drink differentially lowered alcohol consumption, consistent with other data in this species and elsewhere implicating dopamine in the rewarding effects of alcohol. Received: 14 January 1997 / Final version: 10 September 1997     

Sapropterin dihydrochloride, 6-R-L-erythro-5,6,7,8-tetrahydrobiopterin,in the treatment of phenylketonuria

Sapropterin dihydrochloride, 6-R-L-erythro-5,6,7,8-tetrahydrobiopterin (BH4) is being introduced in the US for treatment of phenylketonuria (PKU). This compound has been in use in Europe to treat mild forms of PKU. Tetrahydrobiopterin is the cofactor in the hydroxylation reaction of the three aromatic amino acids phenylalanine, tyrosine and tryptophan. It is also involved in other reactions, which are not the focus of this review. The cofactor BH4 is synthesized in many tissues in the body. The pathway of BH4 biosynthesis is complex, and begins with guanosine triphosphate (GTP). The first reaction that commits GTP to form pterins is GTP cyclohydrolase. Several reactions follow resulting in the active cofactor BH4. During the hydroxylation reaction BH4 is oxidized to quinonoid-BH2, which is recycled by dihydropteridine reductase, resulting in the active cofactor. It was discovered that some patients with PKU had a decline in blood phenylalanine after oral intake of BH4. This response to BH4 is not the result of change in the synthesis or regeneration of the cofactor, but rather an effect on the mutant enzyme phenylalanine hydroxylase either by accommodating the higher K_m of the mutant enzyme or by acting as a chaperone for the mutant enzyme. This response has become of intense interest in the treatment of PKU.     

利福霉素B产生菌─—地中海诺卡氏菌的推理选育

利福霉素Ｂ的生物合成受芳香族氨基酸（Ｔｒｐ、Ｔｙｒ、Ｐｈｅ）的反馈抑制。用紫外线处理Ｎ．ｍｅｄｉｔｅｒｒａｎｅｉＸＣ－１０２菌丝悬浮液后，并在含０．１％芳香族氨基酸的琼脂平板上分离芳香族氨基酸抗性变种。结果表明，芳香族氨基酸抗性变种的发酵效价高于自然分离株和紫外线诱变株；色氨酸抗性变种（Ｔｒｐｒ）的发酵效价又高于酪氨酸抗性变种（Ｔｙｒｒ）和苯丙氨酸抗性变种（Ｐｈｅｒ）；０．５％Ｔｒｐｒ变种比０．１％Ｔｒｐｒ变种更高产。其中０．５％Ｔｒｐｒ变种ＸＣ－５４０的生产能力较出发菌株ＸＣ－１０２增加４２．１６％。传代试验表明ＸＣ－５４０的遗传特性稳定，在７ｍ３罐做发酵放大，与出发菌株相比，发酵效价和发酵指数分别提高５１．１１％和５１．０２％。     

Simultaneous depletion of tryptophan, tyrosine and phenylalanine as an experimental method to probe brain monoamine function in humans

Brain monoamines are important regulators of affective and cognitive processes and are involved in the aetiology of a number of psychiatric disorders. While methods to probe serotonin and catecholamine function are established, limited methods are available to probe monoamine function as a whole in humans. In the current study, we examined if simultaneous depletion of monoamine precursors can be used as a possible probe of monoamine function. Ten healthy subjects were tested under two treatment conditions; balanced control (B) condition and combined monoamine depletion (CMD) condition. Monoamine precursor depletion was associated with significant reductions in plasma-free tryptophan (46%), tyrosine (74%) and phenylalanine (78%). Greater reductions were achieved for ratios of each precursor to other large neutral amino acids (LNAA); tryptophan/LNAA (86%), tyrosine/LNAA (94%) and phenylalanine/LNAA (94%). Findings suggest that simultaneous depletion of monoamine precursors can achieve significant plasma monoamine depletion in the range expected to affect brain monoamine function.