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1.
The aromatic amino acid hydroxylases tyrosine and phenylalanine hydroxylase both contain non-heme iron, utilize oxygen and tetrahydrobiopterin, and are tetramers of identical subunits. The catalytic domains of these enzymes are homologous, and recent X-ray crystallographic analyses show the active sites of the two enzymes are very similar. The hydroxyl oxygens of tyrosine 371 in tyrosine hydroxylase and of tyrosine 325 of phenylalanine hydroxylase are 5 and 4.5 A, respectively, away from the active site iron in the enzymes. To determine whether this residue has a role in the catalytic mechanism as previously suggested [Erlandsen, H., et al. (1997) Nat. Struct. Biol. 4, 995-1000], tyrosine 371 of tyrosine hydroxylase was altered to phenylalanine by site-directed mutagenesis. The Y371F protein was fully active in tyrosine hydroxylation, eliminating an essential mechanistic role for this residue. There was no change in the product distribution seen with phenylalanine or 4-methylphenylalanine as a substrate, suggesting that the reactivity of the hydroxylating intermediate was unaffected. However, the KM value for phenylalanine was decreased 10-fold in the mutant protein. These results are interpreted as an indication of greater conformational flexibility in the active site of the mutant protein. 相似文献
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F Fusetti H Erlandsen T Flatmark RC Stevens 《Canadian Metallurgical Quarterly》1998,273(27):16962-16967
Phenylalanine hydroxylase (PheOH) catalyzes the conversion of L-phenylalanine to L-tyrosine, the rate-limiting step in the oxidative degradation of phenylalanine. Mutations in the human PheOH gene cause phenylketonuria, a common autosomal recessive metabolic disorder that in untreated patients often results in varying degrees of mental retardation. We have determined the crystal structure of human PheOH (residues 118-452). The enzyme crystallizes as a tetramer with each monomer consisting of a catalytic and a tetramerization domain. The tetramerization domain is characterized by the presence of a domain swapping arm that interacts with the other monomers forming an antiparallel coiled-coil. The structure is the first report of a tetrameric PheOH and displays an overall architecture similar to that of the functionally related tyrosine hydroxylase. In contrast to the tyrosine hydroxylase tetramer structure, a very pronounced asymmetry is observed in the phenylalanine hydroxylase, caused by the occurrence of two alternate conformations in the hinge region that leads to the coiled-coil helix. Examination of the mutations causing PKU shows that some of the most frequent mutations are located at the interface of the catalytic and tetramerization domains. Their effects on the structural and cellular stability of the enzyme are discussed. 相似文献
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Extensive restriction site polymorphism at the human phenylalanine hydroxylase locus and application in prenatal diagnosis of phenylketonuria 总被引:4,自引:0,他引:4
AS Lidsky FD Ledley AG DiLella SC Kwok SP Daiger KJ Robson SL Woo 《Canadian Metallurgical Quarterly》1985,37(4):619-634
A total of 10 restriction site polymorphisms have been identified at the human phenylalanine hydroxylase locus using a full-length human phenylalanine hydroxylase cDNA clone as a hybridization probe to analyze human genomic DNA. These polymorphic patterns segregate in a Mendelian fashion and concordantly with the disease state in various PKU kindreds. The frequencies of the restriction site polymorphisms at the human phenylalanine hydroxylase locus among Caucasians are such that the observed heterozygosity in the population is 87.5%. Thus, most families with a history of classical phenylketonuria can take advantage of the genetic analysis for prenatal diagnosis and carrier detection of the hereditary disorder. 相似文献
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T Popescu M Blazkova L Kozak G Jebeleanu A Popescu 《Canadian Metallurgical Quarterly》1998,12(5):314-319
PURPOSE: Simultaneous fluorescence in situ hybridization (FISH) was used in a preimplantation genetic diagnosis program to determine which embryos were normal for chromosomes X, Y, 13, 18, and 21. METHODS: Single blastomeres were disrupted and attached to glass slides using acetic acid and ethanol. Using a ratio mixture of chromosome enumeration DNA probes in combination with locus-specific identifier DNA probes, FISH was performed for the identification of chromosomes X, Y, 13, 18, and 21. RESULTS: Fourteen couples enrolled in IVF produced 134 embryos for biopsy. Blastomeres subjected to five-color FISH revealed that 22% of embryos were normal for chromosomes X, Y, 13, 18, and 21. In addition, 52% were abnormal and no results could be detected for 25%. Twelve couples underwent embryo transfer, two couples did not receive embryos due to lack of any normal embryos, and three couples became pregnant. CONCLUSIONS: The simultaneous detection of five-color FISH is a feasible method to detect aneuploidy in preimplantation embryos from women of advanced maternal age. 相似文献
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Exon 7 of the phenylalanine hydroxylase (PAH) gene was analyzed in 45 children affected with classic phenylketonuria (PKU) from northern China by using PCR-single strand conformation polymorphism (PCR-SSCP) technique and DNA direct sequencing. Six missense mutations (i.e. R243Q, R241H, G247V, L249H, P254I and G257V) and one silent mutation (V245V) were identified. The latter three missense mutations were demonstrated as novel mutations in comparison with the PAH mutation Database. One missense mutation (R241H) was first documented in Chinese. Our results showed population and regional differences in the PAH mutation distribution and suggest that there is more than one founding population for PKU in China. The finding of novel mutations will enhance our capability in molecular diagnosis of PKU. 相似文献
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By using a three-dimensional computed tomography (CT) scanner, we compared the anatomic features of the pelvis of three fetuses of same gestational age, one with a normal pelvis representing the reference model, one with classic bladder exstrophy, and one with cloacal exstrophy. The tomography slices were selected at the same levels for each case. Three angles expressing external opening of the pelvis were defined. Comparing normal and abnormal pelvises allowed definition of three criteria for the correction of the malformation: (a) the sum of the differential angles gives the amplitude of the correction needed; (b) a supraacetabular osteotomy appears to allow best closure of the pelvic ring; (c) only three slices of a CT scan are needed, which cannot be harmful, especially for neonates. Therefore, we believe that a CT scan of the pelvis should be performed whenever an osteotomy is planned in the surgical reconstruction of bladder and cloacal exstrophy. 相似文献
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Two tests have been compared for detection of heterozygotes for phenylketonuria, one based on determination of plasma phenylalanine and tyrosine concentrations in fasting individuals and the other on kinetic evaluation of the plasma elimination curve after intravenous loading with L-phenylalanine. The plasma elimination curve was biexponential and the kinetics were evaluated according to the two-compartment model. The constant, beta, expressing the rate of elimination from plasma at pseudo-equilibrium, the rate constant for the elimination from the central compartment, and the total body clearance were determined. Of these three, total body clearance, which on the average was reduced by 32% in the phenylketonuric heterozygotes, showed the best discriminatory ability, but was not better than the information on concentrations of phenylalanine and tyrosine in detecting heterozygotes for phenylketonuria. 相似文献
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The activity of rat liver phenylalanine hydroxylase (PAH; phenylalanine 4-monooxygenase, EC 1.14.16.1) is regulated by interaction with its substrate, phenylalanine, and its coenzyme, BH4 [tetrahydrobiopterin (6R-dihydroxypropyl-L-erythro-5,6,7,8-tetrahydropterin)]. The structural changes accompanying these interactions have been studied by radiation target analysis. PAH purified from rat liver was incubated with 2 mM phenylalanine to achieve complete activation of the enzyme. Frozen samples were irradiated with various doses of high energy electrons; samples were subsequently thawed, and several surviving properties of the enzyme were determined. Each parameter decreased as a single exponential function of radiation dose. Radiation target analysis of enzymatic activity yielded a dimeric target size. Similar radiation effects on subunit monomers and on tetrameric structure were observed. Together with results from unactivated enzyme, these data show that phenylalanine increases the interactions between the subunits in a dimer and weakens the interactions between dimers in a tetramer. These alterations prevent the natural cofactor, a tetrahydrobiopterin, from exerting a negative effect on activity. 相似文献
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N Hovdenak 《Canadian Metallurgical Quarterly》1980,15(4):401-404
Peroral jejunal biopsies were performed in 63 adult patients from a population of 40,000. Twenty-two patients had definite pathologic biopsy findings; 20 of these had gluten-induced enteropathy. With the 15 previously known patients in the region, this makes a minimum prevalence of 1 per 1,143. Subtotal villous atrophy was found in 8 patients, whereas partial villous atrophy was found in 12 patients. The clinical picture varied considerably, borborygmia, diarrhoea, abdominal pain, and psychiatric symptoms being the most commonly encountered symptoms. Steatorrhoea was found in only two patients with subtotal villous atrophy. In all the remaining patients, including six with subtotal villous atrophy, the faecal fat excretion was normal. A low whole blood folic acid concentration was the most consistent laboratory finding, although some patients had values slightly above the lower normal limit. Anaemia was not pronounced. Iron deficiency, as measured by the erythrocyte protoporphyrin IX concentration, was not common. A high degree of clinical awareness and broad indications for jejunal biopsy-taking is recommended. 相似文献
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As many as 3033 patients with myocardial infarction residing in rural areas were kept under observation. In this population, myocardial reinfarction (MRI) was diagnosed in 411 subjects, with 78.8% having had it for 4 years. Those MRI patients ranging between 51 to 60 years showed the greatest prevalence (44.3%). The ratio of micro- to macrofocal (through-and-through) MRI was 1:3, that of males to females 9:1. Microfocal MRI was commonly associated with a pain-free variant of the disease (23.5%) with low frequency of thromboembolic complications (3.1%). Every fifth patient with macrofocal (through-and-through) renecrosis presented with aneurysm of the heart. In a 10-year and longer follow-up, mortality from MRI was 43.6 percent among patients with macrofocal MRI, while deaths attributable to microfocal MRI were estimated to be 28.9 percent. High mortality rates suggest great severity of illness and low efficacy of the drug therapy adopted. 相似文献
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Neck sprain, a common sequela of motor vehicle trauma, is characterised by soft tissue lesions in the facet joints (the capsule), cervical discs (the annulus), ligaments, muscles, and sometimes in the nerve roots or the central nervous system. The cardinal manifestation of whiplash injury is neck pain, which may arise from any innervated part of the injured joints or muscles except the disc nucleus, the articular surface of facet joints, or the flaval ligaments. Pain may be referred to the skin or muscles via the respective sensory nerves. The tissue lesions, together with post-traumatic nerve root degeneration or medullary compression, may give rise to the complex neck sprain syndrome. Localisation of the source of symptoms by means of meticulous physical examination, MRI (magnetic resonance imaging), or accurately sited local anaesthetic injections is described in the article, and operative and multimodal rehabilitation procedures are discussed. 相似文献
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M Krokowski J Bodalski A Bratek P Machejko S Caillat-Zucman 《Canadian Metallurgical Quarterly》1998,24(3):241-243
Six cancer resection specimens were thoroughly sectioned and microscopically examined at areas known to have been around 21 gauge fine needle aspiration (FNA) biopsy sites, in an attempt to identify needle tracks. All cases had an interval of not less than 10 days between FNA biopsy and surgery. Foci of tumour were identified histologically in needle tracks from two patients with carcinoma. This is the first instance, outside of experimental animal models, of histologically confirmed, viable tumour spread in FNA biopsy tracks. Although this complication is not common and is of unknown clinical significance, it is one that all clinicians who undertake FNA of malignant neoplasms should be aware of. 相似文献
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The clinical study comprising 254 patients with bacterial meningoencephalitis (meningococcal, pneumococcal, staphylococcal, undefined, with n = 135, 76, 43, 120 respectively) permitted identifying a syndrome of oculomotor disturbances. It has been shown that assessment of changes in oculomotor disturbances enables the extent of inflammatory process, focal lesion as well as course and outcome of bacterial meningoencephalitis to be determined in a timely fashion. 相似文献
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Although the importance of the father for the development of the child is well known and although an increased presence of the father in the family system is often demanded, processes of active shutting out the father occur. The deterioration of relation between father and child is not seen as a result of pathological male personality, but rather as a result of family or couple system. The special situation for counsellors of such cases is discussed. 相似文献