Crystal structure of tyrosine hydroxylase at 2.3 Å and its implications for inherited neurodegenerative diseases

Goodwill, Kenneth E.; Sabatier, Christelle; Marks, Cara; Raag, Reetta; Fitzpatrick, Paul F.; Stevens, Raymond C.

doi:10.1038/nsb0797-578

Cited by 279 publications

(359 citation statements)

References 34 publications

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“…Therefore, we hypothesize that the positive charges of the Arg3'-Arg3' sequence contribute to the accessibility of dopamine to or to the expulsion of dopamine from the iron positioned in the active site, as the N-terminus is expected to be localized near the active site (Goodwill et al, 1997). We did not expect that dopamine would bind more to del-38 than to del-35 in the absence of a competitor (as the case of preincubation of the enzyme with dopamine) (Fig.…”

Section: Discussionmentioning

confidence: 99%

Dopamine Inhibition of Human Tyrosine Hydroxylase Type 1 Is Controlled by the Specific Portion in the N‐Terminus of the Enzyme

Nakashima¹,

Mori²,

Suzuki³

et al. 1999

Journal of Neurochemistry

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Tyrosine hydroxylase (TH), which converts L-tyrosine to L-DOPA, is a rate-limiting enzyme in the biosynthesis of catecholamines; its activity is regulated by feedback inhibition by catecholamine products including dopamine. To investigate the specific portion of the N-terminus of TH that determines the efficiency of dopamine inhibition, wild-type and N-terminal 3 5 , 38-, and 44-amino acid-deleted mutants (del-35, del-38, and del-44, respectively) of human TH type l were expressed as a maltose binding protein fusion in Escherichia coli and purified as a tetrameric form by affinity and size-exclusion chromatography. The fused-form wild-type enzyme possessed almost the same specific enzymatic activity as the previously reported recombinant nonfused form. Although maximum velocities of all N-terminus-deleted forms were about one-fourth of the wild-type value, there was no difference in Michaelis constants for L-tyrosine or (6R)-(~-erythro-l',2'-dihydroxypropyl)-2-amino-4-hydroxy-5,6,7,8-tetrahydropteridine (GRBPH,) among the four enzymes. The iron contents incorporated into the three N-terminus-deleted mutants were significantly lower than that of wild type. However, there was no substantial difference in incorporated iron contents among the three mutants. The deletion of up to no less than 38 amino acid residues in the N-terminus made the enzyme more resistant to dopamine inhibition than the wild-type or del-35 TH form. Dopamine bound to the del-38 more than to the del-35 TH form. However, when incubation with dopamine was followed by further inhibition with the cofactor GRBPH, dopamine was expelled more readily from the del-38 than from the del-35 TH form. These observations suggest that the amino acid sequence G l~~~-A r g~~-A r g~~ plays a key role in determining the competition between dopamine and GRBPH, and affects the efficiency of dopamine inhibition of the catalytic activity. Key Words: Tyrosine hydroxylase [TH; tyrosine 3-monooxygenase; L-tyrosine, tetrahydr0pterin:oxygen oxidoreductase (3-hydroxylating); EC 1.14.16.21, which catalyzes the conversion of L-DOPA from L-tyrosine (Nagatsu et al., 1964), is a rate-limiting enzyme in the biosynthesis of catecholamines (Levitt et al., 1965). The enzyme requires a reduced pterin (Nagatsu et al., 1964), and (6R)-(~-erythro-1 ,2'-dihydroxypropyl)-2-amino-4-hydroxy-5,6,7,8-tebxhydropteridine (6R-tebxhydrobiopteri~ 6RBPH4) is the natural cofactor (Kaufman, 1963;Brenneman and Kaufman, 1964;Matsuura et al., 1985). TH also requires ferrous iron (Nagatsu et al., 1964;Shiman et al., 1971). TH consists of a catalytic domain (C-domain) and a regulatory domain (R-domain) (Abate and Joh, 1991). The C-domain is located at the C-terminal two-thirds of the molecule and binds the substrates (L-tyrosine and molecular oxygen) and the cofactor (6RBPH,). In contrast, the R-domain has been assigned to the N-terminal end (Hoeldtke and Kaufman, 1977;Abate et al., 1988;Abate and Joh, 1991) and has an important role in substrate specificity and in control of the catalytic activ...

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Section: Discussionmentioning

confidence: 99%

Dopamine Inhibition of Human Tyrosine Hydroxylase Type 1 Is Controlled by the Specific Portion in the N‐Terminus of the Enzyme

Nakashima¹,

Mori²,

Suzuki³

et al. 1999

Journal of Neurochemistry

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“…We have recently determined the crystal structure of a truncation mutant of rat PAH (lacking 24 C-terminal residues) with catalytic and regulatory properties of the wild-type protein. 11 Previously, the crystal structures of smaller fragments, lacking the regulatory domain, of human PAH 12,13 and a homologous enzyme, rat tyrosine hydroxylases (TyrOH) 14 have also been determined. Jointly, this structural information now allows a structural interpretation of any PAH mutation resulting in PAH deficiency.…”

Section: Introductionmentioning

confidence: 99%

“…Similarly, structural interpretations of selected mutations in PAH have been reported recently. 4,[12][13][14]22 However, the criteria for selection of mutations analysed in these studies have not been detailed, and the effects have been discussed without the structural knowledge of the entire protein. To gain more general insights into the molecular basis of PAH deficiency, we chose in this study to correlate the metabolic phenotype and expression studies with the structural knowledge for a larger unbiased set of mutations.…”

Section: Introductionmentioning

confidence: 99%

Structural interpretation of mutations in phenylalanine hydroxylase protein aids in identifying genotype–phenotype correlations in phenylketonuria

Jennings

Cotton²,

Kobe

2000

Eur J Hum Genet

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Phenylalanine hydroxylase (PAH) is the enzyme that converts phenylalanine to tyrosine as a rate-limiting step in phenylalanine catabolism and protein and neurotransmitter biosynthesis. Over 300 mutations have been identified in the gene encoding PAH that result in a deficient enzyme activity and lead to the disorders hyperphenylalaninaemia and phenylketonuria. The determination of the crystal structure of PAH now allows the determination of the structural basis of mutations resulting in PAH deficiency. We present an analysis of the structural basis of 120 mutations with a 'classified' biochemical phenotype and/or available in vitro expression data. We find that the mutations can be grouped into five structural categories, based on the distinct expected structural and functional effects of the mutations in each category. Missense mutations and small amino acid deletions are found in three categories: 'active site mutations', 'dimer interface mutations', and 'domain structure mutations'. Nonsense mutations and splicing mutations form the category of 'proteins with truncations and large deletions'. The final category, 'fusion proteins', is caused by frameshift mutations. We show that the structural information helps formulate some rules that will help predict the likely effects of unclassified and newly discovered mutations: proteins with truncations and large deletions, fusion proteins and active site mutations generally cause severe phenotypes; domain structure mutations and dimer interface mutations spread over a range of phenotypes, but domain structure mutations in the catalytic domain are more likely to be severe than domain structure mutations in the regulatory domain or dimer interface mutations. European Journal of Human Genetics (2000) 8, 683-696.

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“…The similar structures of the catalytic domains [9][10][11] and a variety of mechanistic studies suggest that the three aromatic amino acid hydroxylases share a common catalytic mechanism [12]. For all three enzymes, hydroxylation of the physiological substrate occurs via an electrophilic aromatic substitution reaction with an activated oxygen species [13][14][15].…”

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confidence: 97%

“…The reactivities of the hydroxylating intermediates in all three enzymes are similar [17] and they have overlapping substrate specificities [18][19][20], supporting the proposal for a common Fe(IV)O intermediate. The iron atom in each is coordinated by two histidines and a glutamate [9][10][11]21,22]. This metal-binding arrangement has been named a "2-His-1-carboxylate facial triad" and is also found in a variety of nonheme iron enzymes with different three-dimensional structures, including the extradiol and Rieske dioxygenases, the α-ketoglutarate dependent hydroxylases, and isopenicillin N synthase [23].…”

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confidence: 99%

Characterization of metal ligand mutants of phenylalanine hydroxylase: Insights into the plasticity of a 2-histidine-1-carboxylate triad

Fitzpatrick

2008

Archives of Biochemistry and Biophysics

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The iron atom in the nonheme iron monooxygenase phenylalanine hydroxylase is bound on one face by His285, His290, and Glu330. This arrangement of metal ligands is conserved in the other aromatic amino acid hydroxylases, tyrosine hydroxylase and tryptophan hydroxylase. A similar 2-His-1-carboxylate facial triad of two histidines and an acidic residue are the ligands to the iron in other nonheme iron enzymes, including the α-ketoglutarate dependent hydroxylases and the extradiol dioxygenases. Previous studies of the effects of conservative mutations of the iron ligands in tyrosine hydroxylase established that there is some plasticity in the nature of the ligands and that the three ligands differ in their sensitivity to mutagenesis. To determine the generality of this finding for enzymes containing a 2-His-1-carboxylate facial triad, the His285, His290, and Glu330 in rat phenylalanine hydroxylase were mutated to glutamine, glutamate, and histidine. All of the mutant proteins had low but measurable activities for tyrosine formation. In general, mutation of Glu330 had the greatest effect on activity and mutation of His290 the least. All of the mutations resulted in an excess of tetrahydropterin oxidized relative to tyrosine formation, with mutation of His285 having the greatest effect on the coupling of the two partial reactions. The H285Q enzyme had the highest activity as tetrahydropterin oxidase at 20% the wild-type value. All of the mutations greatly decreased the affinity for iron, with mutation of Glu330 the most deleterious. The results complement previous results with tyrosine hydroxylase in establishing the plasticity of the individual iron ligands in this enzyme family.Phenylalanine hydroxylase (PheH) 1 is a tetrahydropterin-dependent nonheme enzyme that catalyzes the hydroxylation of phenylalanine to tyrosine, an important pathway for phenylalanine catabolism (Scheme 1) [1]. As a liver enzyme, PheH is critical for catabolizing excess phenylalanine in the diet. Consequently, a deficiency in PheH results in the debilitating disease phenylketonuria [2]. PheH belongs to the small family of aromatic amino acid hydroxylases, all of which use tetrahydrobiopterin as the source of electrons to hydroxylate the aromatic ring of an aromatic amino acid. Tyrosine hydroxylase (TyrH) and tryptophan †

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Crystal structure of tyrosine hydroxylase at 2.3 Å and its implications for inherited neurodegenerative diseases

Cited by 279 publications

References 34 publications

Dopamine Inhibition of Human Tyrosine Hydroxylase Type 1 Is Controlled by the Specific Portion in the N‐Terminus of the Enzyme

Dopamine Inhibition of Human Tyrosine Hydroxylase Type 1 Is Controlled by the Specific Portion in the N‐Terminus of the Enzyme

Structural interpretation of mutations in phenylalanine hydroxylase protein aids in identifying genotype–phenotype correlations in phenylketonuria

Characterization of metal ligand mutants of phenylalanine hydroxylase: Insights into the plasticity of a 2-histidine-1-carboxylate triad

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