Erythrocyte catechol-O-methyltransferase and plasma dopamine-β-hydroxylase in human umbilical cord blood

Summary Plasma dopamine--hydroxylase enzymatic activity and immunoreactive protein levels in human umbilical cord blood are only about 2% as great as values in the blood of older subjects. Erythrocyte catechol-O-methyltransferase activity levels in umbilical cord blood are very similar to those in the blood of adult subjects.This work was supported in part by NIH grants NS 11014 and HL 17487-1. Dr Weinshilboum is an Established Investigator of the American Heart Association. We thank Luanne Wussow and the physicians and nurses of the Department of Obstetrics and Gynecology of the Mayo Clinic for their assitance with these studies.     

Elevation of plasma dopamine-β-hydroxylase activity during insulin-induced hypoglycemia in man

Résumé Nous avons étudié l'activité de la dopamine--hydroxylase du plasma humain. Elle a été remarquablement accure par l'administration de grandes quantités d'insuline. Les corrélations entre cette activité et celle des systèmes nerveux sympathiques sont également discutées.     

Correlation between serum dopamine-β-hydroxylase activity and dopamine-β-hydroxylase and tyrosine hydroxylase activities in central and peripheral adrenergic neurons and adrenal glands

Summary Serum dopamine--hydroxylase activity in spontaneously hypertensive rats and Wistar-Kyoto rats had a positive correlation with dopamine--hydroxylase and tyrosine hydroxylase activities in mesenteric vessels, vas deferens, and adrenal glands at 14–16 weeks of age, a negative correlation with dopamine--hydroxylase activity in locus coeruleus at 3 weeks and 14–16 weeks of age, and a positive correlation with tyrosine hydroxylase activity only at 3 weeks of age, but not at 14–16 weeks of age.     

Adrenal dopamine-β-hydroxylase activity: 24-hour rhythmicity and evidence for pineal control

Summary Adrenal medullary dopamine--hydroxylase activity was found in male rats to have a 24-hour rhythm, with an approximately 6-fold increase at about the time of the onset of darkness. This nocturnal rise in enzyme activity did not occur when lights were kept on, nor did it occur in animals that had been pinealectomized.Supported in part by grants from the Ford Foundation (No. 630-05050A), National Institutes of Health (No. HD-03352) and National Institute of Mental Health (No. MH-25019) USPHS.     

A radioimmunoassay for dopamine-β-hydroxylase using a glass-bound antibody

Résumé Les auteurs décrivent une méthode radioimmunologique de dosage de la DH utilisant un anticorps lié de façon covalente à des billes de verre. La technique a été appliquée à la mesure de la DH dans les surrénales et différentes régions du cerveau de buf. Les principaux avantages de cette méthode sont sa relative facilité d'emploi, sa précision et sa haute spécificité.

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This work was supported by USPHS Grants Nos. MH-02717, NS-06801 and CA-02071 and NSF Grant No. GB-27603.Acknowledgements: The authors wish to express their appreciation for the technical assistance of Susan Kadner and William P. Vann.     

Changes in dopamineβ-hydroxylase activity of monkey plasma with age

Summary Plasma dopamine -hydroxylase activity of Japanese monkeys (Macaca fuscata fuscata) increased with age, and the developmental changes were similar to those of human beings. However, the adult level plasma DBH activity of various monkey species was much lower than that of human beings.We thank Dr.K. Nozawa, Dr.O. Takenaka and Mr.T. Shotake (Primate Research Institute, Kyoto University) for their help in collecting monkey blood samples.     

Human Genome and Diseases:¶Dyskeratosis congenita

Dyskeratosis congenita is an inherited skin and bone marrow failure syndrome. There are X-linked, autosomal dominant and autosomal recessive forms of the disease. The X-linked form is due to mutations in the DKC1 gene at Xq28. The encoded protein, dyskerin, is a component of both small nucleolar ribonuclear protein particles and the telomerase complex. Mutations in DKC1 mainly lead to amino acid substitutions. The autosomal dominant form of the disease is due to mutations in hTR, the RNA component of telomerase, making it likely that the disease is due to defective telomerase activity. Mutations in hTR are predicted to either disrupt secondary structure or alter the template region. The gene or genes involved in the recessive forms of the disease remain elusive, though genes whose products are required for telomere maintenance are strong candidates.     

Human Genome and Diseases:¶Double-strand breaks and translocations in cancer

The correct repair of double-strand breaks (DSBs) is essential for the genomic integrity of a cell, as inappropriate repair can lead to chromosomal rearrangements such as translocations. In many hematologic cancers and sarcomas, translocations are the etiological factor in tumorigenesis, resulting in either the deregulation of a proto-oncogene or the expression of a fusion protein with transforming properties. Mammalian cells are able to repair DSBs by pathways involving homologous recombination and nonhomologous end-joining. The analysis of translocation breakpoints in a number of cancers and the development of model translocation systems are beginning to shed light on specific DSB repair pathway(s) responsible for the improper repair of broken chromosomes. Received 19 June 2001; received after revision 6 September 2001; accepted 11 September 2001     

Kinetic studies on soluble and membrane-bound dopamineβ-hydroxylase isolated from storage vesicles of heart and adrenal medulla of different species

Summary IdenticalK _m-values for soluble and membrane-bound dopamine -hydroxylase isolated from adrenal medullary vesicles of different species were obtained; the same holds true for both forms of the enzyme of heart vesicles.The excellent technical assistance of MissSabine Klemt is gratefully acknowledged.     

Human and rodent type 1 11β-hydroxysteroid dehydrogenases are 7β-hydroxycholesterol dehydrogenases involved in oxysterol metabolism

Interconversion between cortisone and the glucocorticoid receptor ligand cortisol is carried out by 11-hydroxysteroid dehydrogenase (11-HSD)isozymes and constitutes a medically important example of pre-receptor control of steroid hormones. The enzyme 11-HSD type 1 (11-HSD1) catalyzes the conversion of cortisone to its active receptor-binding derivative cortisol, whereas 11-HSD type 2 performs the reverse reaction. Specific inhibitors against the type 1 enzyme lower intracellular levels of glucocorticoid hormone, with an important clinical application in insulin resistance and other metabolic disorders. We report here on the in vitro oxysterol-metabolizing properties of human and rodent 11-HSD1. The enzyme, either as full-length, membrane-attached, or as a transmembrane domain-deleted, soluble form, mediates exclusively conversion between 7-ketocholesterol and 7-hydroxycholesterol with similar k_cat values as observed with glucocorticoid hormones. Thus, human, rat, and mouse 11-HSD1 have dual enzyme activities like the recently described 7-hydroxysteroid dehydrogenase/11-hydroxysteroid dehydrogenase from hamster liver, but differ fundamentally from the latter in that 7-OH rather than 7-OH dehydrogenase constitutes the second activity. These results demonstrate an enzymatic origin of species differences in 7-oxysterol metabolism, establish the origin of endogenous 7-OH cholesterol in humans, and point to a possible involvement of 11-HSD1 in atherosclerosis.Received 30 December 2003; received after revision 16 February 2004; accepted 16 February 2004     

Human Genome and Diseases:¶Cellular and molecular aspects of Zellweger syndrome and other peroxisome biogenesis disorders

Peroxisomes are single-membrane-bound organelles present in virtually all eukaryotic cells. They are involved in numerous metabolic processes, both catabolic and anabolic, including β-oxidation of very long chain fatty acids, metabolism of hydrogen peroxide, plasmalogen biosynthesis and bile acid synthesis. In several genetic diseases, there is either isolated deficiency of a specific peroxisomal protein (single-protein deficiencies) or a defect in the formation of the organelle with loss of multiple peroxisomal functions (peroxisome biogenesis disorders). X-linked adrenoleukodystrophy is an example of the former, and the Zellweger spectrum of the latter. Peroxisome biogenesis disorders are inherited in an autosomal recessive manner and result from mutations in any of at least 12 PEX genes that encode peroxins. This article reviews the peroxisomal system, the clinical, biochemical and molecular aspects of peroxisomal disorders, and discusses recent scientific advances in the understanding of peroxisome biogenesis. Received 16 October 2001; received after revision 2 January 2002; accepted 3 January 2002     

Human Genome and Diseases:¶The molecular pathogenesis of the Marfan syndrome

The Marfan syndrome (MFS) is an autosomal dominant heritable disorder of connective tissue with highly variable clinical manifestations including aortic dilatation and dissection, ectopia lentis, and a range of skeletal anomalies. Mutations in the gene for fibrillin-1 (FBN1) cause MFS and other related disorders of connective tissue collectively termed type-1 fibrillinopathies. Fibrillin-1 is a main component of the 10- to 12-nm extracellular microfibrils that are important for elastogenesis, elasticity, and homeostasis of elastic fibers. Mutations in fibrillin-1 are hypothesized to exert their effects by dominant negative mechanisms, but recent work has also emphasized the potential role of proteases and disturbances in tissue homeostasis in the pathogenesis of the MFS. This article provides an overview of the clinical aspects of the MFS and current thinking on the pathogenesis of this disorder.     

Human Genome and Diseases:¶Apaf1 in developmental apoptosis and cancer: how many ways to die?

Apaf1 has been described as the core of the apoptosome. Deficiency in murine Apaf1 leads to embryonic lethality with a phenotype affecting many aspects of developmental apoptosis. In the developing brain, Apaf1 is a death regulator of the neuronal founder cells. Combined intercrosses of mouse lines mutant for members of the mitochondrial death pathway are providing us with some clues about the relative regulation existing among neuronal cell populations. Apaf1-deficient embryos display an interesting phenotype in the inner ear and in limb development, which involves different caspase-dependent and -independent pathways. Moreover, APAF1 is mutated in human melanomas, and its depletion contributes to malignant transformation in a mouse model of cancer. This review has a double aim: the analysis of the alternatives taken by the embryo to bring into the suicidal program different cells at different stages, and the relevance of APAF1 in the onset and progression of cancer. Received 5 March 2001; received after revision 19 April 2001; accepted 4 May 2001     

Completion of the natural groups of ergot alkaloids: Syntheses and pharmacological profiles ofβ-ergosine andβ-ergoptine

Summary The syntheses and pharmacological potencies of -ergosine and -ergoptine, the missing links in the natural groups of ergot peptide alkaloids are described.86th communication on ergot alkaloids.     

Dopamine-β-hydroxylase,adrenaline, noradrenaline and dopamine in the venous blood of adrenal gland of man: A comparison with levels in the periphery of the circulation

Summary In 10 human subjects plasma dopamine--hydroxylase activity was found in the adrenal vein blood to be as high as in the periphery of the circulation. Adrenaline concentration in the adrenal vein blood was in the mean 170 times, noradrenaline concentration 11 times and dopamine concentration little higher than levels in the periphery.This study was supported by the Deutsche Forschungsgemeinschaft.     

Palmitoylation by DHHC3 is critical for the function, expression, and stability of integrin α6β4

The laminin-binding integrin α6β4 plays key roles in both normal epithelial and endothelial cells and during tumor cell progression, metastasis, and angiogenesis. Previous cysteine mutagenesis studies have suggested that palmitoylation of α6β4 protein supports a few integrin-dependent functions and molecular associations. Here we took another approach and obtained strikingly different results. We used overexpression and RNAi knockdown in multiple cell types to identify protein acyl transferase DHHC3 as the enzyme responsible for integrin β4 and α6 palmitoylation. Ablation of DHHC3 markedly diminished integrin-dependent cellular cable formation on Matrigel, integrin signaling through Src, and β4 phosphorylation on key diagnostic amino acids (S1356 and 1424). However, unexpectedly, and in sharp contrast to prior α6β4 mutagenesis results, knockdown of DHHC3 accelerated the degradation of α6β4, likely due to an increase in endosomal exposure to cathepsin D. When proteolytic degradation was inhibited (by Pepstatin A), rescued α6β4 accumulated intracellularly, but was unable to reach the cell surface. DHHC3 ablation effects were strongly selective for α6β4. Cell-surface levels of ~10 other proteins (including α3β1) were not diminished, and the appearance of hundreds of other palmitoylated proteins was not altered. Results obtained here demonstrate a new substrate for the DHHC3 enzyme and provide novel opportunities for modulating α6β4 expression, distribution, and function.